An overview of chelicerate ovaries, with special reference to mites - myths and facts.

In arthropods of the subphylum Chelicerata a panoistic ovary, in which all germline cells differentiate into oocytes, prevails. Among the chelicerates, mites are believed to show a great variety of the structure of the female gonads. In general, the knowledge of the ovarian structure in mites is fragmentary and patchy. In both evolutionary lines, Acariformes and Parasitiformes, apart from the panoistic ovary, the meroistic ovary, in which the oocytes grow supported by their sibling cells, the nurse cells, occurs. The presence of the meroistic ovary is considered an apomorphic state. Previous studies revealed a various structure of the meroistic ovary in different mite taxa, and the differences came down, inter alia, to a different number and location of the nurse cells in relation to the oocytes. Here we provide a comprehensive review of the structure of the Chelicerata ovary, with special reference to the mite ovary. We also provide our preliminary results of the analysis of ovarian structure in two representatives of terrestrial Parasitengona (Acariformes), Allothrombium fuliginosum (Trombidiidae) and Erythraeus cinereus (Erythraeidae), performed using light, confocal and electron transmission microscopy. The analyses allowed for verification of data published before. In A. fuliginosum we showed the presence of the nurse cells in the ovarian wall, so the ovary should be classified as meroistic. In meroistic ovary of E. cinereus we found that each oocyte is connected to several mononucleated nurse cells. The verification of literature data and broadening the knowledge of the structure of the female gonad in mites, will result in estimating the usefulness of the ovary traits in phylogenetic analyses and will provide the basis for inference about the directions of evolutionary changes of female gonad at lower systematic levels.

Spectacular alterations in the female reproductive system during the ovarian cycle and adaptations for matrotrophy in chernetid pseudoscorpions (Pseudoscorpiones: Chernetidae).

Pseudoscorpions are small matrotrophic chelicerates. The embryos develop in a brood sac and feed on the nutritive fluid provided by the female. It was widely accepted that the nutritive fluid is synthesized in the ovary. Recent studies have shown that in Chelifer cancroides, a representative of Cheliferidae, considered one of the most derived pseudoscorpion families, the nutritive fluid is produced not only in the ovary but also in the oviducts. Since evolution of adaptations for matrotrophy in pseudoscorpions is poorly known, we aimed to verify our hypothesis that pseudoscorpions of the family Chernetidae, closely related to Cheliferidae, share the traits of adaptations to matrotrophy in the structure and function of the female reproductive system with C. cancroides. We analysed the structure of the ovary and oviducts in five representatives of chernetids with light, confocal, and transmission electron microscopy. The results confirmed our hypothesis and provided new data which broaden our knowledge of matrotrophic pseudoscorpions. We show that in chernetids, the ovary and oviducts undergo significant alterations including their size, multistep hypertrophy and polyploidization of the epithelial cells involved in secretion of the nutritive fluid, the complex secretory activity of the epithelial cells, massive degeneration of the epithelial cells that have completed secretion, and epithelium renewal.

Small body size of pseudoscorpions and a distinct architecture of the ovary: A step to miniaturization?

Chelicerata, the second largest subphylum of Arthropoda, includes invertebrates with a wide range of body size. Pseudoscorpions are among small or miniature chelicerates which exhibit several morphological, anatomical, and developmental features related to miniaturization, e.g., replacement of book lungs by tracheae, unpaired gonads, and matrotrophic development of the embryos outside the female body, in the brood sac. In this paper, we show the ovary structure of two pseudoscorpion species, Cheiridium museorum and Apocheiridium ferum (Cheiridiidae). Both cheiridiids are one of the smallest pseudoscorpions. The results of our observations conducted in light, transmission electron, and confocal microscopy demonstrate that the ovary of C. museorum and A. ferum, displays a significant structural difference that is unusual for chelicerates. The difference concerns the spatially restricted position of the germarium. We show that such ovary architecture results in a significantly reduced number of growing oocytes and in consequence a reduced number of deposited eggs. A centrally located germarium implies also a modified pattern of ovary development during oocyte growth due to long distance migration of the germline and the accompanying somatic cells. Herein, we postulate that such an ovary structure is related to the pseudoscorpion's small body size and it is a step towards miniaturization in the smaller pseudoscorpions species.

Adaptations for matrotrophy in the female reproductive system in the pseudoscorpion Chelifer cancroides (Chelicerata: Pseudoscorpiones, Cheliferidae).

Pseudoscorpiones (pseudoscorpions, false scorpions) is an order of small terrestrial chelicerates. While most chelicerates are lecithotrophic, that is, embryos develop due to nutrients (mostly yolk) deposited in the oocyte cytoplasm, pseudoscorpions are matrotrophic, that is, embryos are nourished by the female. Pseudoscorpion oocytes contain only a small amount of yolk. The embryos develop within a brood sac carried on the abdominal site of the female and absorb nutrients by a pumping organ. It is believed that in pseudoscorpions nutrients for developing embryos are produced in the ovary during a postovulatory (secretory) phase of the ovarian cycle. The goal of our study was to analyze the structure of the female reproductive system during the secretory phase in the pseudoscorpion Chelifer cancroides, a representative of the family Cheliferidae, considered to be one of the most advanced pseudoscorpion taxa. We use diverse microscopic techniques to document that the nutritive fluid is produced not only in the ovaries but also by the epithelial cells in the oviducts. The secretory active epithelial cells are hypertrophic and polyploid and release their content by fragmentation of apical parts. Our observations also indicate that fertilization occurs in the oviducts. Moreover, in contrast to previous findings, we show that secretion of the nutritive material starts when the fertilized oocytes reach the brood sac and thus precedes formation of the pumping organ. Summing up, we show that C. cancroides exhibits traits of advanced adaptations for matrotrophy due to coordinated secretion of the nutritive fluid by the ovarian and oviductal epithelial cells, which substantially increases the efficiency of nutritive fluid formation. Since the secretion of nutrients starts before formation of the pumping organ, we suggest that the embryos are able to absorb the nutritive fluid also in the early embryonic stages.

Morphology of Ovaries and Oogenesis in Chelicerates.

The subphylum Chelicerata represents one of the oldest groups among arthropods and comprises more than a dozen orders. Representatives of particular orders differ significantly in their external morphology, reproductive biology, behavior, and structure of internal organs, e.g. of the respiratory system. However, in almost all chelicerates (excluding some mites) the female gonads show a similar architecture. In this chapter, the chelicerate-type ovary structure and the course of oogenesis are described. Structural and functional diversities of the chelicerate-type ovary in non-matrotrophic and matrotrophic arachnids are also presented.

Differentiation of follicular epithelium in polytrophic ovaries of Pieris napi (Lepidoptera: Pieridae)-how far to Drosophila model.

Lepidoptera together with its sister group Trichoptera belongs to the superorder Amphiesmenoptera, which is closely related to the Antliophora, comprising Diptera, Siphonaptera, and Mecoptera. In the lepidopteran Pieris napi, a representative of the family Pieridae, the ovaries typical of butterflies are polytrophic and consist of structural ovarian units termed ovarioles. Each ovariole is composed of a terminal filament, germarium, vitellarium, and ovariole stalk. The germarium houses developing germ cell clusters and somatic prefollicular and follicular cells. The significantly elongated vitellarium contains linearly arranged ovarian follicles in successive stages of oogenesis (previtellogenesis, vitellogenesis, and choriogenesis). Each follicle consists of an oocyte and seven nurse cells surrounded by follicular epithelium. During oogenesis, follicular cells diversify into five morphologically and functionally distinct subpopulations: (1) main body follicular cells (mbFC), (2) stretched cells (stFC), (3) posterior terminal cells (pFC), (4) centripetal cells (cpFC), and (5) interfollicular stalk cells (IFS). Centripetal cells are migratorily active and finally form the micropyle. Interfollicular stalk cells derive from mbFC as a result of mbFC intercalation. Differentiation and diversification of follicular cells in Pieris significantly differ from those described in Drosophila in the number of subpopulations and their origin and function during oogenesis.

Formation of the diverticular lumen that enables oocyte fertilization in katoikogenic scorpions, Heterometrus spinifer and Opistophthalmus boehmi (Scorpionidae).

The structure of diverticula of the female gonads was analyzed in two scorpions from the family Scorpionidae by means of standard microscopic techniques (light microscopy, histochemistry, transmission electron microscopy). In scorpions, the female gonad, termed the ovariuterus, participates in two consecutive processes: oogenesis and embryogenesis. In sexually reproducing scorpions, the ovariuterus is also involved in fertilization. Both scorpions under study reproduce sexually. They also represent the katoikogenic type of development, which means that oogenesis, fertilization, and embryogenesis take place in the diverticula, which are sac-like outpocketings of the ovariuterine tubules. Formation of a lumen in the diverticulum is indispensable for sperm entry to enable fertilization and subsequent embryogenesis. The aim of the study was to test our hypothesis that the diverticulum lumen forms due to the engagement of the centrally located stalk cells. In this report, we show that in two species of katoikogenic scorpions, at the final stages of oogenesis, the cytoplasm of the stalk cells contains secretory organelles. In the stages preceding fertilization, secretory activity of the stalk cells and fragmentation of their apical parts lead to formation of the diverticulum lumen by a process similar to "cord hollowing" that commonly occurs for lumen formation in other morphogenesis model systems.

Scorpion katoikogenic ovariuterus - Much more alike to apoikogenic type than it seemed to be.

Scorpions are viviparous matrotrophic arthropods. Both, fertilization and embryonic development occur in the female gonad called ovariuterus. Two distinct reproductive patterns are recognized among scorpions: apoikogenic and katoikogenic. In the ovariuterus of apoikogenic scorpions growing oocytes protrude from the ovarian wall and continue previtellogenic and vitellogenic growth on the gonad surface being accompanied by the follicular cells that cover the oocyte surface, and, in most families, the stalk cells that join the oocyte with the ovariuterus wall. In the katoikogenic ovariuterus the oocytes grow in outpocketings of the ovarian wall called diverticula. The aim of our study was to show the development and structure of the diverticula in two katoikogenic scorpions from the family Scorpionidae: Ophistothalmus boehmei and Heterometrus spinifer. We show that the somatic components of each diverticulum develop from the two epithelial layers of the ovariuterine wall. Before fertilization, the wall of the mature diverticula consists of two distinctive epithelial layers: an internal and an external one. Our observations reveal that the epithelial cells of the internal layer of the diverticulum show striking morphological resemblance to the follicular and stalk cells that accompany the growing oocytes in some apoikogenic scorpions. The external epithelial layer of the katoikogenic diverticulum seems to have no equivalents in the apoikogenic type. Functions of the somatic cells of the diverticulum are discussed.

Asymmetry in structure of the eggshell in Osmylus fulvicephalus (Neuroptera: Osmylidae): an exceptional case of breaking symmetry during neuropteran oogenesis.

Ovaries of neuropterans are of meroistic-polytrophic type. The ovarian tubes, the ovarioles, are divided into two major parts: a germarium, comprised of newly formed germ cell clusters; and a vitellarium, housing linearly arranged ovarian follicles. Each ovarian follicle consists of the germ cell cluster diversified into different number of nurse cells, and the oocyte enclosed by follicular epithelium. In Osmylus fulvicephalus, a representative of Neuroptera, during consecutive stages of oogenesis, the follicular cells undergo a multistep process of diversification which leads to the appearance of several follicular cell subpopulations i.e., the main-body follicular cells, the stretched cells, the anterior centripetal cells, and posterior centripetal cells. The anterior centripetal cells occupy the anterior pole of the oocyte and in advanced oogenesis due to hypertrophy that transform into anterior fold cells. Initially, the anterior fold cells form a symmetric fold, but in advanced oogenesis, quite different from other neuropterans studied so far, they undergo uneven hypertrophic growth which results in breaking symmetry of the anterior fold that becomes shifted to the ventral side of the oocyte. Since the anterior fold cells participate in the production of the specialized chorion structure, the micropyle, asymmetric structure of the anterior fold, is reflected both in its asymmetric position and in the asymmetric construction of the micropyle. As a consequence of breaking symmetry of the anterior fold, Osmylus eggshell gains dorso-ventral polarity, which is unusual for neuropterans.

Ultrastructural analysis of the ovary and oogenesis in Spinicaudata and Laevicaudata (Branchiopoda) and its phylogenetic implications.

Recent molecular studies have indicated a close relationship between Crustacea and Hexapoda and postulated their unification into the Pancrustacea/Tetraconata clade. Certain molecular analyses have also suggested that the crustacean lineage, which includes the Branchiopoda, might be the sister group of Hexapoda. We test this hypothesis by analyzing the structure of the ovary and the ultrastructural features of oogenesis in two branchiopod species, Cyzicus tetracerus and Lynceus brachyurus, representing two separate orders, Spinicaudata and Laevicaudata, respectively. The female gonads of these species have not been investigated before. Here, we demonstrate that in both studied species the ovarian follicles develop inside characteristic ovarian protrusions and comprise a germline cyst surrounded by a simple somatic (follicular) epithelium, supported by a thin basal lamina. Each germline cyst consists of one oocyte and three supporting nurse cells, and the oocyte differentiates relatively late during ovarian follicle development. The synthesis of oocyte reserve materials involves rough endoplasmic reticulum and Golgi complexes. The follicular cells are penetrated by a complex canal system and there is no external epithelial sheath covering the ovarian follicles. The structure of the ovary and the ultrastructural characteristics of oogenesis are not only remarkably similar in both Cyzicus and Lynceus, but also share morphological similarities with Notostraca as well as the basal hexapods Campodeina and Collembola. Possible phylogenetic implications of these findings are discussed.

Balbiani body, nuage and sponge bodies--term plasm pathway players.

In many animal species, germ cells are specified by maternally provided, often asymmetrically localized germ cell determinant, termed the germ plasm. It has been shown that in model organisms such as Xenopus laevis, Danio rerio and Drosophila melanogaster germ plasm components (various proteins, mRNAs and mitochondria) are delivered to the proper position within the egg cell by germline specific organelles, i.e. Balbiani bodies, nuage accumulations and/or sponge bodies. In the present article, we review the current knowledge on morphology, molecular composition and functioning of these organelles in main lineages of arthropods and different ovary types on the backdrop of data derived from the studies of the model vertebrate species.

Differentiation of somatic cells in the ovariuteri of the apoikogenic scorpion Euscorpius italicus (Chelicerata, Scorpiones, Euscorpiidae).

In apoikogenic scorpions, growing oocytes protrude from the gonad (ovariuterus) and develop in follicles exposed to the mesosomal (i.e. hemocoelic) cavity. During subsequent stages of oogenesis (previtellogenesis and vitellogenesis), the follicles are connected to the gonad surface by prominent somatic stalks. The aim of our study was to analyze the origin, structure and functioning of somatic cells accompanying protruding oocytes. We show that these cells differentiate into two morphologically distinct subpopulations: the follicular cells and stalk cells. The follicular cells gather on the hemocoelic (i.e. facing the hemocoel) surface of the oocyte, where they constitute a cuboidal epithelium. The arrangement of the follicular cells on the oocyte surface is not uniform; moreover, the actin cytoskeleton of these cells undergoes significant modifications during oocyte growth. During initial stages of the stalk formation the stalk cells elongate and form F-actin rich cytoplasmic processes by which the stalk cells are tightly connected to each other. Additionally, the stalk cells develop microvilli directed towards the growing oocyte. Our findings indicate that the follicular cells covering hemocoelic surfaces of the oocyte and the stalk cells represent two distinct subpopulations of epithelial cells, which differ in morphology, behavior and function.

Differentiation and function of the ovarian somatic cells in the pseudoscorpion, Chelifer cancroides (Linnaeus, 1761) (Chelicerata: Arachnida: Pseudoscorpionida).

Pseudoscorpion females carry fertilized eggs and embryos in specialized brood sacs, where embryos are fed with a nutritive fluid produced and secreted by somatic ovarian cells. We used various microscopic techniques to analyze the organization of the somatic cells in the ovary of a pseudoscorpion, Chelifer cancroides. In young specimens, the ovary is a cylindrical mass of internally located germline cells (oogonia and early previtellogenic oocytes) and two types of somatic cells: the epithelial cells of the ovarian wall and the internal interstitial cells. In subsequent stages of the ovary development, the oocytes grow and protrude from the ovary into the hemocoel (opisthosomal cavity). At the same time the interstitial cells differentiate into the follicular cells that directly cover the oocyte surface, whereas some epithelial cells of the ovarian wall form the oocyte stalks - tubular structures that connect the oocytes with the ovarian tube. The follicular cells do not seem to participate in oogenesis. In contrast, the cells of the stalk presumably have a dual function. During ovulation the stalk cells appear to contribute to the formation of the external egg envelope (chorion), while in the post-ovulatory phase of ovary function they cooperate with the other cells of the ovarian wall in the production of the nutritive fluid for the developing embryos.

Reptilian myotomal myogenesis-lessons from the sand lizard Lacerta agilis L. (Reptilia, Lacertidae)Update.

Reptilian myotomal myogenesis is poorly understood. This paper reports on structural, ultrastructural and immunocytochemical studies of muscle differentiation in sand lizard (Lacerta agilis) embryos. During somitogenesis, the somites are composed of epithelial vesicles with a centrally located somitocoel. At later developmental stages the ventral portion of the somite cortex disaggregates into the sclerotome mesenchyme, while the dorsal wall of the somite differentiates into dermomyotome. At these developmental stages, mononucleated cells of the dermomyotome are Pax3-positive. The dermomyotome layer forms the dorsomedial and ventromedial lips. The myotome is first composed of mono- and then of multinucleated myotubes and small mononucleated cells that occur in the vicinity of the myotubes. These mononucleated cells exhibit low proliferative potential as revealed by the use of PCNA antibody. At subsequent stages of myogenesis the mononucleated cells express Pax7 protein, a marker of satellite cells, and assume ultrastructural features characteristic of satellite cells. Some of the mononucleated cells contribute to muscle growth, being involved in fusion with differentiating muscle fibers. This study revealed similarities of myotomal myogenesis in reptiles to that of other vertebrates.

How is myogenesis initiated in chordates?

Expression of transcriptor factors MyoD, Myf5, myogenin and MRF4 forms the basis of myogenesis. In Acrania, Pisces and Amphibia, as in Aves, myogenesis is initiated by MyoD. In Mammalia expression of Myf5 initiates myogenesis. Signal proteins Wnt and Shh induce the expression of genes encoding for MyoD or Myf5. In fishes and amphibians expression of MyoD starts in non-segmented mesoderm and then in myotomal cells. In birds and mammals expression of MyoD or Myf5 is initiated in the cells of the dermatomyotome. Embryonic myotomes are post-mitotic. Proliferating cells Pax3 and Pax7-positive and mesenchymal cells take part in the growth of myotomal muscles. Cells migrating to the limb bud contain regulatory proteins Six4/Six1, Pax3, Lbx1 and c-met. Rectus abdominis develops from cells that contain Pax3 and Lbx1.

Differences in the relative timing of developmental events during oogenesis in lower dipterans (Nematocera) reveal the autonomy of follicular cells' differentiation program.

Although the ovaries of Nematocera are of the same meroistic-polytrophic type, they show significant differences in the activity of germ cells (oocytes, nurse cells) and their relative contribution to ribosome synthesis and storage during oogenesis. These different activities result in the different growth rate of the germ cells and may determine the life span of the nurse cells. Comparative analysis revealed that with reference to germ cell activity, two basic types of oogenesis in Nematocera can be distinguished. In the Tinearia type, the nurse cells grow considerably and are active until advanced stages of oogenesis, whereas the oocyte is transcriptionally inert. Conversely, in the Tipula type of oogenesis, the oocyte nucleus contains transcriptionally active multiple nucleoli, while nurse cells probably do not contribute to ribosome synthesis, remain relatively small and degenerate early in oogenesis. We studied and compared the process of somatic follicular cell differentiation in nematoceran species representing both types of oogenesis. Our observations indicate that morphogenesis of the follicular cells is at least partly independent of the nurse cell activity, while the execution of their differentiation does not require direct contacts between the follicular cells and the oocyte.

Yolk nucleus--the complex assemblage of cytoskeleton and ER is a site of lipid droplet formation in spider oocytes.

Oocytes (future egg cells) of various animal groups often contain complex organelle assemblages (Balbiani bodies, yolk nuclei). The molecular composition and function of Balbiani bodies, such as those found in the oocytes of Xenopus laevis, have been recently recognized. In contrast, the functional significance of more complex and highly ordered yolk nuclei has not been elucidated to date. In this report we describe the structure, cytochemical content and evolution of the yolk nucleus in the oocytes of a common spider, Clubiona sp. We show that the yolk nucleus is a spherical, rather compact and persistent cytoplasmic accumulation of several different organelles. It consists predominantly of a highly elaborate cytoskeletal scaffold of condensed filamentous actin and a dense meshwork of intermediate-sized filaments. The yolk nucleus also comprises cisterns of endoplasmic reticulum, mitochondria, lipid droplets and other organelles. Nascent lipid droplets are regularly found in the cortical regions of the yolk nucleus in association with the endoplasmic reticulum. Single lipid droplets become surrounded by filamentous cages formed by intermediate filaments. Coexistence of the forming lipid droplets with the endoplasmic reticulum in the cortical zone of the yolk nucleus and their later investment by intermediate-sized filamentous cages suggest that the yolk nucleus is the birthplace of lipid droplets.

The Balbiani body in the oocytes of a common cellar spider, Pholcus phalangioides (Araneae: Pholcidae).

Previtellogenic oocytes of a common cellar spider, Pholcus phalangioides, contain a single aggregation of organelles referred here to as the Balbiani body. It is a well defined ooplasmic structure predominantly composed of fine granular nuage, RNA rich material but comprising also mitochondria, vesicles of endoplasmic reticulum and stacks of Golgi cysternae. The Balbiani body originates early during previtellogenesis in the form of a cap-shaped mass in juxtaposition to one pole of the oocyte nucleus. During later stages of previtellogenic growth the Balbiani body translocates as a single body towards the ooplasm periphery. The results presented indicate that Balbiani body translocation is cytoskeleton independent. Balbiani body repositioning does not result in the localization of its components to any distinct, asymmetrically situated region of the ooplasm but, instead, ends up with their even dispersion in the oocyte cortex. The Balbiani body in Pholcus does not seem to be implicated either in germ cell determination or organelle inheritance. Its homology with similar organelle accumulations in the oocytes of other species is discussed.

Ovariole development in telotrophic ovaries of snake flies (Raphidioptera).

Snake flies (Raphidioptera), alder flies (Megaloptera: Sialidae) and also some myxophagan coleopterans share the same, peculiar telotrophic organization of their ovarioles usually referred to as ovarioles of the Sialis-type. Ovariole ontogenesis in Raphidia sp. is described and the basic events that lead to the formation of germ cell clusters and their subsequent transformations are reported. It was found that the major cellular events during ovariole formation in Raphidia and Sialis are essentially the same. Discrepancies concern details of germ cell cluster formation, differentiation of cystocytes within clusters and their location within the developing tropharium. Based on these results the hypothetical model of the Sialis-type ovariole formation, previously presented by King and Büoning (1985) is verified. A hypothesis on the mechanisms of oocyte determination in telotrophic ovaries is also presented.

Formation and structure of nutritive cords in telotrophic ovarioles of snake flies (Insecta: Raphidioptera).

Telotrophic ovariole of Raphidia spp. is composed of the anteriorly located terminal filament, tube-shaped tropharium, the vitellarium and the ovariole stalk. The tropharium consists of a central syncytial core surrounded by one cell thick layer of tapetum cells. Early previtellogenic oocytes differentiate at the base of tropharium. Both the oocytes and the tapetum cells are connected with the central syncytium by delicate intercellular bridges. At the onset of previtellogenic growth, the anterior parts of the oocytes become extended and form long cytoplasmic projections--nutritive cords. Each nutritive cord contains numerous microtubules that show no preferential orientation within the cord but diminishing anterior-posterior gradient of distribution. Irregular arrangement of microtubules indicates that this cytoskeletal scaffold does not play any role in directed transport within the ovariole but instead constitutes one of the elements of the structural framework of the nutritive cord. Besides microtubules, the stability of the nutritive cords in Raphidia ovarioles is maintained by the rim-shaped membrane foldings lined with microfilaments.