Izabela Jędrzejowska

Balbiani body, nuage and sponge bodies - The germ 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.

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.

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.

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.

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.

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

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.

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.

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.

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.

On the histology, ultrastructure and function of the spermoviduct and free oviduct in egg-retaining door snails (Pulmonata: Clausiliidae)

We studied the anatomical, histological and ultrastructural changes of the spermoviduct and free oviduct in three closely related clausiliid snails (Baleinae) in relation to their different reproductive strategies: oviparity in Laciniaria plicata, egg retention in Vestia gulo and viviparity in Alinda biplicata. Since egg retention occurs in the lower parts of the spermoviduct and free oviduct, these organs were studied in detail to determine their structural diversity. The results of anatomical studies showed that the length proportions of the free oviduct and spermoviduct are different between oviparous snails and egg-retaining/viviparous snails. The allospermiduct of the free oviduct displays significant morphological and ultrastructural dissimilarities in snails that differ in reproductive strategies. These differences mainly concern the type of subepithelial secretory cells and their secretory products. In egg-retaining snails, the subepithelial cells are serous and their secretory products create a suitable environment (oncotic pressure) for retained eggs, whereas in oviparous snails the serous cells are absent.