Ovary architecture of two branchiobdellid species and Acanthobdella peledina (Annelida, Clitellata)

The aim of this study was to present data about ovary organization and oogenesis in two small groups of clitellate annelids, i.e. in representatives of Acanthobdellida (Acanthobdella peledina) and Branchiobdellida (Branchiobdella pentodonta and Branchiobdella parasitica), and to compare them to ovaries known from true leeches and oligochaetous clitellates. In A. peledina, the ovaries have the form of elongated cords, termed ovary cords, and are enveloped by coelomic sacs, the so-called ovisacs. The ovisacs are paired and each one contains only one ovary cord. The morphology and structure of the ovary cords depend on the maturity level of the animal. In young specimens the ovary cords are short and contain mainly oogonial cells and germ cells entering meiosis. Oogonia divide mitotically without full cytokineses, and as a result germ-line cysts are formed. As the animals grow, the cords become more elongated and the germ cells within the cords differentiate into nurse cells and oocytes. Oocytes gather cell organelles and, finally, detach from the ovary cord and float freely in the ovisac lumen.In both examined branchiobdellidans the ovaries are also paired. They are short and conical and are not enclosed within ovisacs. The narrow end of each ovary is connected to the intersegmental septum via a ligament, whereas the outermost (broad) end of the ovary extends freely into the coelom. The ovaries are polarized. Their narrow ends contain oogonia, whereas nurse cells and growing oocytes, gradually projecting from the ovary, can be found in their middle and outermost parts. Early vitellogenic oocytes detach from the ovary and float freely in the coelom.In all of the species studied, the ovaries are made up of germ-line cysts associated with somatic (follicular) cells. The architecture of a germ-line cyst is exactly the same as in other clitellate annelids that have been studied to date. Each germ cell in a cyst has one stable cytoplasmic bridge connecting it with a central anuclear cytoplasmic mass, a cytophore. The fate of germ cells constituting cysts is diverse. The majority of the cells withdraw from meiosis and become nurse cells; only a few continue meiosis, grow and become oocytes. The meroistic mode of oogenesis is suggested. We suggest also that the formation of germ-line cysts and ovary meroism should be regarded as basal conditions for all Clitellata. The occurrence of ovisacs enveloping the ovaries in A. peledina and Hirudinida is regarded as a synapomorphy of both groups, whereas ovaries found in B. pentodonta and B. parasitica have no ovisacs and resemble ovaries described in Oligochaeta sensu stricto.     

Ovary organization and oogenesis in two species of Lumbriculida (Annelida,Clitellata)

The aim of the present study is to describe the organization of the ovary and mode of oogenesis at the ultrastructural level in two representatives of Lumbriculida – Lumbriculus variegatus and Stylodrilus heringianus. In both species studied, the ovaries are small and conically shaped structures that are attached to the intersegmental septum via a thin ligament. The ovaries are composed of germline cysts formed by germ cells interconnected by stable cytoplasmic bridges. As a rule, the cyst center is occupied by a poorly developed anuclear cytoplasmic mass, termed a cytophore, whereas the germ cells are located at the periphery of the cyst. Germline cysts are enveloped by somatic cells. The ovaries of the species studied are polarized, i.e., along the long axis of the ovary there is an evident gradient of germ cell development. The data obtained suggest ovary meroism, i.e., two categories of germ cells were found: oocytes, which continue meiosis, gather nutrients, grow and protrude into the body cavity, and nurse cells, which do not grow and are supposed to supply oocytes with cell organelles and macromolecules via the cytophore. The ovary structure and mode of oogenesis in the species studied were compared with those of other clitellate annelids. As a rule, in all clitellates studied to date, the ovaries are composed of germline cysts equipped with a cytophore and associated with somatic cells; however, the ovary morphology differs between taxa regarding several quantitative and qualitative features. The ovary organization and mode of oogenesis in L. variegatus and S. heringianus strongly resemble those found in Tubificinae and Branchiobdellida studied to date. Our results also support a sister-group relationship between Lumbriculida and a clade comprising ectoparasitic clitellates (i.e., Branchiobdellida, Acanthobdellida and Hirudinida) with Branchiobdellida as a plesiomorphic sister group to Acanthobdellida and Hirudinida.     

The Ovary of Tubifex tubifex (Clitellata,Naididae, Tubificinae) Is Composed of One,Huge Germ-Line Cyst that Is Enriched with Cytoskeletal Components

Recent studies on the ovary organization and oogenesis in Tubificinae have revealed that their ovaries are small polarized structures that are composed of germ cells in subsequent stages of oogenesis that are associated with somatic cells. In syncytial cysts, as a rule, each germ cell is connected to the central cytoplasmic mass, the cytophore, via only one stable intercellular bridge (ring canal). In this paper we present detailed data about the composition of germ-line cysts in Tubifex tubifex with special emphasis on the occurrence and distribution of the cytoskeletal elements. Using fixed material and live cell imaging techniques, we found that the entire ovary of T. tubifex is composed of only one, huge multicellular germ-line cyst, which may contain up to 2,600 cells. Its architecture is broadly similar to the cysts that are found in other clitellate annelids, i.e. a common, anuclear cytoplasmic mass in the center of the cyst and germ cells that are connected to it via intercellular bridges. The cytophore in the T. tubifex cyst extends along the long axis of the ovary in the form of elongated and branched cytoplasmic strands. Rhodamine-coupled phalloidin staining revealed that the prominent strands of actin filaments occur inside the cytophore. Similar to the cytophore, F-actin strands are branched and they are especially well developed in the middle and outermost parts of the ovary. Microfilaments are also present in the ring canals that connect the germ cells with the cytophore in the narrow end of the ovary. Using TubulinTracker, we found that the microtubules form a prominent network of loosely and evenly distributed tubules inside the cytophore as well as in every germ cell. The well-developed cytoskeletal elements in T. tubifex ovary seem to ensure the integrity of such a huge germ-line cyst of complex (germ cells - ring canals - cytophore) organization. A comparison between the cysts that are described here and other well-known female germ-line cysts is also made.     

Germ-line cysts are formed during oogenesis in Erpobdella octoculata (Annelida,Clitellata, Erpobdellidae)

Abstract

Erpobdella octoculata (Clitellata, Hirudinea, Erpobdellidae) has paired ovarian sacs, each containing several rod-shaped structures termed ovarian bodies. Oogenesis takes place within the ovarian bodies. We show that in the apical part of the bodies the germ-line cells form syncytial cysts of cells interconnected by stable intercellular bridges. Germ-line cyst architecture is broadly similar to that of other clitellate annelids; that is, each germ cell has only one intercellular bridge connecting it to the anuclear cytoplasmic mass, the cytophore. Unlike germ-line cysts described in other leech species, the cytophore in cysts of E. octoculata is poorly developed, taking the form of thin cytoplasmic strands. Oogenesis in E. octoculata is meroistic because the germ cells forming the cysts (cystocytes) have diverse fates, i.e., nurse cells and oocytes appear. One large ramified cell (apical cell) occurs within the apical part of the ovarian body. We compare the ultrastructure of the apical cell found in E. octoculata with that of apical cells described recently in some hirudiniform leeches. The germ-line cysts as well as the oocytes are enveloped by somatic follicular cells. As in other leeches, the follicular cells surrounding the growing oocytes have cytoplasm perforated by intracellular canals. In view of the many similarities between E. octoculata ovarian bodies and the ovary cords described in glossiphoniids and especially in hirudiniform leeches, we suggest that the ovarian bodies found in E. octoculata are in fact modified ovary cords.     

An ultrastructural study of the ovary cord organization and oogenesis in the amphibian leech Batracobdella algira (Annelida,Clitellata, Hirudinida)

This study reveals the ovary micromorphology and the course of oogenesis in the leech Batracobdella algira (Glossiphoniidae). Using light, fluorescence, and electron microscopies, the paired ovaries were analyzed. At the beginning of the breeding season, the ovaries were small, but as oogenesis progressed, they increased in size significantly, broadened, and elongated. A single convoluted ovary cord was located inside each ovary. The ovary cord was composed of numerous germ cells gathered into syncytial groups, which are called germ-line cysts. During oogenesis, the clustering germ cells differentiated into two functional categories, i.e., nurse cells and oocytes, and therefore, this oogenesis was recognized as being meroistic. As a rule, each clustering germ cell had one connection in the form of a broad cytoplasmic channel (intercellular bridge) that connected it to the cytophore. There was a synchrony in the development of the clustering germ cells in the whole ovary cord. In the immature leeches, the ovary cords contained undifferentiated germ cells exclusively, from which, previtellogenic oocytes and nurse cells differentiated as the breeding season progressed. Only the oocytes grew considerably, gathered nutritive material, and protruded at the ovary cord surface. The vitellogenic oocytes subsequently detached from the cord and filled tightly the ovary sac, while the nurse cells and the cytophore degenerated. Ripe eggs were finally deposited into the cocoons. A comparison of the ovary structure and oogenesis revealed that almost all of the features that are described in the studied species were similar to those that are known from other representatives of Glossiphoniidae, which indicates their evolutionary conservatism within this family.

     

Ovary cords organization in Hirudo troctina Johnson, 1816 and Limnatis nilotica (Savigny, 1822) (Clitellata,Hirudinea)

In both examined species of Hirudinea there are paired spheroid ovisacs, and within each ovisac two convoluted ovary cords occur. The morphology of the cords is characteristic: their apical end is club-shaped, the central part is narrow and may contain developing oocytes, whereas the basal end of the cord is irregularly shaped and composed of degenerating cells. The ovary cords are built of somatic and germ-line cells; the latter are united into syncytial cysts. Each germ cell in such a cyst has only one stable cytoplasmic bridge connecting it to the central anuclear cytoplasmic mass, the cytophore. Initially all germ-line cells in a given cyst are morphologically identical, then the fates of cells diversify. Most of them become nurse cells and eventually degenerate; the rest continue meiosis, gather macromolecules, cell organelles and nutritive material and become oocytes. The oogenesis found in the species studied should be regarded as meroistic. Previtellogenic oocytes protrude from the cord into the ovisac lumen, whereas the vitellogenic ones float freely in the ovisac lumen. The somatic cells found in the ovary cords are: follicular cells which form the envelope of the cord and are also found among germ cells inside the cord, and one, huge apical cell that always is located at the top of the club-shaped end of the ovary cord. The apical cell has several characteristic features, e.g., it forms long cytoplasmic projections filled with intermediate filaments and it is connected to the neighbouring cells (both somatic and germ-line) via hemidesmosomes. We suggest that the apical cell forms the niche for maintaining germ and somatic stem cells. Generally, the organization of the ovary cords found in both studied species is broadly similar to those described in other hirudiniform leeches studied to date.     

Microorganization of ovaries and oogenesis of Haplotaxis sp. (Clitellata: Haplotaxidae)

Ovaries of Haplotaxis sp. were studied in active and nonactive states, that is, in a sexually mature specimen and in specimens outside of the reproductive period. Two pairs of ovaries were found in segments XI and XII. Especially in the nonactive state, they were in close contact with copulatory glands. Each ovary was composed of germ cells interconnected with syncytial cysts, which were enveloped by a layer of somatic cells. Within cysts each germ cell had one ring canal connecting it to the common anuclear cytoplasmic mass called a cytophore. During oogenesis clustering germ cells differentiated into nurse cells and oocytes; thus, the oogenesis was recognized as meroistic. Vitellogenic oocytes were detached from the ovaries and continued yolk absorption within the body cavity. Because recent studies have shown the variety of ovaries and germ line cyst organization in clitellates and suggest their evolutionary conservatism at the family or subfamily level, the data presented here can be valid in understanding the phylogenetic relationships among Clitellata. In this context, ovaries found in Haplotaxis sp. resembled those of the “Tubifex” type. “Tubifex” ovaries are characteristic for numerous microdrile taxa (tubificines, limnodriloidines, propappids, lumbriculids, and leech‐like branchiobdellids) and can be regarded as the primary character for these Clitellata in which germ‐line cysts are formed during early oogenesis. As the family Haplotaxidae is currently considered to be paraphyletic and the species studied here belongs to Haplotaxidae sensu stricto, our results support the close relationship of Haplotaxidae sensu stricto to the clade consisting of Lumbriculidae, Branchiobdellida, and Hirudinida, in which lumbriculids are sister to the latter two.     

Ovary cord structure and oogenesis in Hirudo medicinalis and Haemopis sanguisuga (Clitellata, Annelida): remarks on different ovaries organization in Hirudinea

In Hirudo medicinalis and Haemopis sanguisuga, two convoluted ovary cords are found within each ovary. Each ovary cord is a polarized structure composed of germ cells (oogonia, developing oocytes, nurse cells) and somatic cells (apical cell, follicular cells). One end of the ovary cord is club-shaped and comprises one huge apical cell, numerous oogonia, and small cysts (clusters) of interconnected germ cells. The main part of the cord contains fully developed cysts composed of numerous nurse cells connected via intercellular bridges with the cytophore, which in turn is connected by a cytoplasmic bridge with the growing oocyte. The opposite end of the cord degenerates. Cord integrity is ensured by flattened follicular cells enveloping the cord; moreover, inside the cord, some follicular cells (internal follicular cells) are distributed among germ cells. As oogenesis progresses, the growing oocytes gradually protrude into the ovary lumen; as a result, fully developed oocytes arrested in meiotic metaphase I float freely in the ovary lumen. This paper describes the successive stages of oogenesis of H. medicinalis in detail. Ovary organization in Hirudinea was classified within four different types: non-polarized ovary cords were found in glossiphoniids, egg follicles were described in piscicolids, ovarian bodies were found characteristic for erpobdellids, and polarized ovary cords in hirudiniforms. Ovaries with polarized structures equipped with apical cell (i.e. polarized ovary cords and ovarian bodies) (as found in arhynchobdellids) are considered as primary for Hirudinea while non-polarized ovary cords and the occurrence of egg follicles (rhynchobdellids) represent derived condition.     

Oogenesis in four species of Piscicola (Hirudinea, Rhynchobdellida)

Piscicola has a pair of elongated sac-shaped ovaries. Inside the ovaries are numerous small somatic cells and regularly spherical egg follicles. Each follicle is composed of three types of cells: many (average 30) germ cells (cystocytes) interconnected by intercellular bridges in clones (cysts), one intermediate cell, and three to five outer follicle cells (envelope cells). Each germ cell in a clone has one intercellular bridge connecting it to the central anucleate cytoplasmic mass, the cytophore. Each cluster of germ cells is completely embedded inside a single huge somatic follicle cell, the intermediate (interstitial) cell. The most spectacular feature of the intermediate cell is its development of a system of intracytoplasmic canals apparently formed of invaginations of its cell membrane. Initially the complex of germ cell cluster + intermediate cell is enclosed within an envelope composed of squamous cells. As oogenesis progresses the envelope cells gradually degenerate. All the germ cells that have terminated their mitotic divisions are of similar size and enter meiotic prophase, but one of the cystocytes promptly starts to grow faster and differentiates into the oocyte, whereas the remaining cystocytes withdraw from meiosis and become nurse cells (trophocytes). Numerous mitochondria, ER, and a vast amount of ribosomes are transferred from the trophocytes via the cytophore toward the oocyte. Eventually the oocyte ingests all the content of the cytophore, and the trophocytes degenerate. Little vitellogenesis takes place; the oocyte gathers nutrients in the form of small lipid droplets. At the end of oogenesis, an electron-dense fibrous vitelline envelope appears around the oocyte, among short microvilli. At the same time, electron-dense cortical granules occur in the oocyte cortical cytoplasm; at the end of oogenesis they are numerous, but after fertilization they disappear from the ooplasm. In the present article we point out many differences in the course of oogenesis in two related families of rhynchobdellids: piscicolids and glossiphoniids.     

Barbronia weberi (Clitellata,Hirudinida, Salifidae) has ovary cords of the Erpobdella type

The organization of the ovaries in representative of the Salifidae (Hirudinida, Erpobdelliformes) was studied at the ultrastructural level for the first time. Like in other leeches, the ovaries of Barbronia weberi are composed of an outer envelope (i.e., an ovisac made up of two coelomic epithelia, muscle cells, and connective tissue) and several internal units, which are broadly similar to the ovary cords found in representatives of the Erpobdellidae. There are usually 6–8 ovary cords that are twisted or cambered with a narrow apical part and a broader, irregularly shaped distal end in each ovisac of B. weberi. Each ovary cord is built from somatic and germ‐line cells and the latter tend to form multicellular cysts that are equipped with a central cytoplasmic core (cytophore). There are two morphologically different subpopulations of germ‐line cells: oocytes and more numerous nurse cells. Growing oocytes form protuberances on the ovary cord surface and eventually detach from the cord and float freely in the ovisac lumen, whereas the other components of germ‐line cysts (i.e., nurse cells and cytophore) degenerate. It should be pointed out that there is a prominent gradient of germ‐cell development along the long axis of the cord. The somatic cells form the ovary cord envelope (the so‐called spongiosa cells) and also penetrate the spaces between germ‐line cells. Both kinds of the somatic cells, that is, those forming the cord envelope and the somatic cells that are associated with oocytes (follicular cells) have a well‐developed system of intercellular channels. Additionally, one prominent somatic cell, the apical cell, was found at the apical tip of each ovary cord. Because the aforementioned features of ovary cords found in B. weberi are very similar (with a few minor exceptions) to the ovary cords that have been described in Erpobdella octoculata and E. johanssoni, we propose the term “ovary cords of the Erpobdella type” for them. Our results support a close phylogenetic relationship between Salifidae and Erpobdellidae. J. Morphol. 275:479–488, 2014. © 2013 Wiley Periodicals, Inc.     

Oogenesis in the leech Glossiphonia heteroclita (Annelida, Hirudinea, Glossiphonidae). I. Ovary structure and previtellogenic growth of oocytes

Glossiphonia heteroclita has paired ovaries whose shape and dimensions change as oogenesis proceeds: during early previtellogenesis they are small and club-shaped, whereas during vitellogenesis they broaden and elongate considerably. During early oogenesis (previtellogenesis), each ovary is composed of an outer envelope (ovisac) that surrounds the ovary cavity and is filled with hemocoelomic fluid, in which a single and very convoluted ovary cord is bathed. The ovary cord consists of germline cells, including nurse cells and young oocytes surrounded by a layer of elongated follicle cells. Additionally, follicle cells with long cytoplasmic projections occur inside the ovary cord, where they separate germ cells from each other. The ovary cord contains thousands of nurse cells. Each nurse cell has one intercellular bridge, connecting it to a central anucleate cytoplasmic mass, the cytophore (rachis); it in turn is connected by one intercellular bridge with each growing oocyte. Numerous mitochondria, RER cisternae, ribosomes, and Golgi complexes are transported from the nurse cells, via the intercellular bridge and cytophore, to the growing oocytes. Oogenesis in G. heteroclita is synchronous with all oocytes in the ovary in the same stage of oogenesis. The youngest observed oocytes are slightly larger than nurse cells, and usually occupy the periphery of the ovary cord. As previtellogenesis proceeds, the oocytes gather a vast amount of cell organelles and become more voluminous. As a result, in late previtellogenesis the oocytes gradually protrude into the ovary cavity. Simultaneously with oocyte growth, the follicle cells differentiate into two subpopulations. The morphology of the follicle cells surrounding the nurse cells and penetrating the ovary cord does not change, whereas those enveloping the growing oocytes become more voluminous. Their plasma membrane invaginates deeply, forming numerous broad vesicles that eventually seem to form channels or conducts through which the hemocoelomic fluid can easily access the growing oocytes.     

Formation of germ-line cysts with a central cytoplasmic core is accompanied by specific orientation of mitotic spindles and partitioning of existing intercellular bridges

Animal germ cells tend to form clonal groups known as clusters or cysts. Germ cells within the cyst (cystocytes) are interconnected by intercellular bridges and thus constitute a syncytium. Our knowledge of the mechanisms that control the formation of germ-cell clusters comes from extensive studies carried on model organisms (Drosophila, Xenopus). Germ-cell clusters have also been described in worms (annelids, flat worms and nematodes), although their architecture differs significantly from that known in arthropods or vertebrates. Their peculiar feature is the presence of a central anucleate cytoplasmic core (cytophore, rachis) around which the cystocytes are clustered. Each cystocyte in such a cluster always has one intercellular bridge connecting it to the central cytoplasmic core. The way that such clusters are formed has remained a riddle for decades. By means of light, fluorescence and electron microscopy, we have analysed the formation and architecture of cystocyte clusters during early stages of spermatogenesis and oogenesis in a few species belonging to clitellate (oligochaetous) annelids. Our data indicate that the appearance of germ cells connected via a central cytophore is accompanied by a specific orientation of the mitotic spindles during cystocyte divisions. Spindle long axes are always oriented tangentially to the surface of the cytophore. In consequence, cystocytes divide perpendicularly to the plane of the existing intercellular bridge. Towards the final stages of cytokinesis, the contractile ring of the cleavage furrow merges with the rim of the intercellular bridge that connects the dividing cystocyte with the cytophore and forces partition of the existing bridge into two new bridges. This work was supported by the following research grants: 2P04C004 28 from the Ministry of Science and Informatization (to P. Świątek and J. Klag) and DS/1018/IZ/2007 (to J. Kubrakiewicz).     

Reductions and new inventions dominate oogenesis of Strepsiptera (Insecta)

The endoparasitic life of strepsipterans (Insecta), especially neotenic females, reduces to a great extent external and internal organs. Light and electron microscopic investigation of ovaries of Elenchus tenuicornis (Kirby) confirms the following: (1) somatic tissues of ovaries are totally reduced, with the exception of some cells surrounding germ cell clusters; (2) a previtellogenic growth phase of oocytes is reduced; (3) nurse cells remain diploid and their membranes degenerate at the onset of vitellogenesis; (4) vitellogenesis is reduced, vitellin and fat vacuoles contribute only 50% to the final egg volume; and (5) chorionogenesis is reduced to a vitellin membrane. However, some features of normal development remain, allowing classification of the ovary type as polytrophic meroistic: (1) germ cells undergo synchronized, incomplete divisions, following the 2ⁿ rule, where all former intercellular bridges become localized in one cystocyte, while the other has none; and (2) only one cell is determined as the oocyte, all other cystocytes serve as nurse cells and the surrounding somatic cells transform into follicular cells. Novel events in oogenesis of strepsipterans include fission of clusters during the phase of cluster mitoses, and protection of oocyte nuclei, while nurse cell nuclei degenerate in the same cytoplasm.     

Ultrastructure and cluster formation in ovaries of bark lice,Peripsocus phaeopterus (Stephens) and Stenopsocus stigmaticus (Imhof and Labram) (Insecta : Psocoptera)

Ultrastructure and previtellogenic growth of ovaries of Peripsocus phaeopterus (Stephens) and Stenopsocus stigmaticus (Imhof and Labram) (Insecta : Psocoptera) are described. The germ cell cluster formation was analyzed in an ovariole of a nymph using ultrathin serial sectioning. Fifteen germ cell clusters were found; 13 contained 4 cystocytes each, while 2 clusters, situated in the very tip, were composed of 2 cystocytes each. A fully developed cluster rises by 2 synchronized mitotic divisions, each followed by incomplete cytokinesis. Microtubules derived from the preceding mitoses form a transient midbody within the intercellular bridge. Later on, a fusome fills each bridge, while at fusomal rims parallel oriented microtubules are tightly associated. Some of these microtubules stretch to cell membranes nearby. The fusomes fuse into a polyfusome and a rosette is thus formed by which all intercellular bridges are drawn together. All cystocytes enter the prophase of meiosis up to pachynema. One of the 2 inner cells continues meiosis and develops as an oocyte, whereas all others transform into nurse cells. After rosette formation, the polyfusome-associated microtubules vanish and some time later, when the nurse cell-oocyte differentiation becomes apparent, the polyfusome itself becomes destroyed. The intercellular bridge, joining the first nurse cell with the 3rd moves away from the other 2. During previtellogenesis, 5 phases can be distinguished, 2 of which are interpreted as logarithmical growth phases with different slopes. The whole set of characters elaborated here for the polytrophic meroistic ovary of psocopterans is fully consistent with the characters of polytrophic meroistic ovaries of Holometabola, indicating a monophyletic origin.     

Early oogenesis in the short-tailed fruit bat Carollia perspicillata: transient germ cell cysts and noncanonical intercellular bridges

The ovaries of early embryos (40 days post coitum/p.c.) of the bat Carollia perspicillata contain numerous germ-line cysts, which are composed of 10 to 12 sister germ cells (cystocytes). Variability in the number of cystocytes within the cyst and between the cysts (defying the Giardina rule) indicates that the mitotic divisions of the cystoblast are asynchronous in this bat species. Serial section analysis showed that the cystocytes are interconnected via intercellular bridges that are atypical, strongly elongated, short-lived, and rich in microtubule bundles and microfilaments. During slightly later stages of embryonic development (44-46 days p.c.), somatic cells penetrate the cyst, and their cytoplasmic projections separate individual oocytes. Separated oocytes surrounded by a single layer of somatic cells constitute the primordial ovarian follicles. The oocytes of C. perspicillata are similar to mouse oocytes and are asymmetric. In both species, this asymmetry is clearly recognizable in the localization of the Golgi complexes. The presence of germ-line cysts and intercellular bridges (although noncanonical) in the fetal ovaries of C. perspicillata suggest that the formation of germ-line cysts is an evolutionarily conserved phase in the development of the female gametes in a substantial part of the animal kingdom.     

Histological studies on early oogenesis in barfin flounder (Verasper moseri)

There is much information on oogenesis from the resumption of the first meiotic division to oocyte maturation in many vertebrates; however, there have been very few studies on early oogenesis from oogonial proliferation to the initiation of meiosis. In the present study, we investigated the histological changes during early oogenesis in barfin flounder (Verasper moseri). In fish with a total length (TL) of 50mm (TL 50mm fish), active oogonial proliferation was observed. In TL 60mm fish, oocytes with synaptonemal complexes were observed. Before the initiation of active oogonial proliferation, somatic cells which surrounded a few oogonial germ cells, started to proliferate to form the oogonial cysts that accompanied oogonial proliferation. In TL 70mm fish, however, the cyst structure of the oocyte was gradually broken by the invagination of somatic cells, and finally the oocyte became a single cell surrounded by follicle cells. Upon comparison of nuclear size, DNA-synthesizing germ cells could be divided into two types: small nuclear cells and large nuclear cells. Based on histological observation, we propose that the small nuclear cells were in the mitotic prophase of oogonia and the large nuclear cells were in the meiotic prophase of oocytes, and that the nuclear size increases upon the initiation of meiosis.     

The larval development of the telotrophic meroistic ovary in the bug Dysdercus intermedius (Heteroptera, Pyrrhocoridae)

Bug ovaries are of the telotrophic meroistic type. Nurse cells are restricted to the anterior tropharium and are in syncytial connection with the oocytes via the acellular trophic core region into which cytoplasmic projections of oocytes and nurse cells open. The origin of intercellular connections in bug ovaries is not well understood. In order to elucidate the cellular processes underlying the emergence of the syncytium, we analysed the development of the ovary of Dysdercus intermedius throughout the five larval instars. Up to the third instar, the germ cell population of an ovariole anlage forms a single, tight rosette. In the center of the rosette, phosphotyrosine containing proteins and f-actin accumulate. This center is filled with fusomal cytoplasm and closely interdigitating cell membranes known as the membrane labyrinth. With the molt to the fourth instar germ cells enhance their mitotic activity considerably. As a rule, germ cells divide asynchronously. Simultaneously, the membrane labyrinth expands and establishes a central column within the growing tropharium. In the fifth instar the membrane labyrinth retracts to an apical position, where it is maintained even in ovarioles of adult females. The former membrane labyrinth in middle and posterior regions of the tropharium is replaced by the central core to which nurse cells and oocytes are syncytially connected. Germ cells in the most anterior part of the tropharium, i.e. those in close proximity to the membrane labyrinth remain proliferative. The posterior-most germ cells enter meiosis and become oocytes. The majority of the ovarioles' germ cells, located in between these two populations, endopolyploidize and function as nurse cells. We conclude that the extensive multiplication of germ cells and their syncytial assembly during larval development is achieved by incomplete cytokineses followed by massive membrane production. Membranes are degraded as soon as the trophic core develops. For comparative reasons, we also undertook a cursory examination of early germ cell development in Dysdercus intermedius males. All results were compatible with the known basic patterns of early insect spermatogenesis. Germ cells run through mitotic and meiotic divisions in synchronous clusters emerging from incomplete cytokineses. During the division phase, the germ cells of an individual cluster are connected by a polyfusome rich in f-actin.     

The telotrophic ovary known from Neuropterida exists also in the myxophagan beetle <Emphasis Type="Italic">Hydroscapha natans</Emphasis>

The ovary structure of the myxophagan beetle, Hycdoscapha natans, was investigated by means of light and electron microscopy for the first time. Each of the two ovaries consists of three ovarioles, the functional units of insect oogenesis. The ovary type is telotrophic meroistic but differs strongly from the telotrophic ovary found among all polyphagous beetles investigated so far. All characters found here are typical of telotrophic ovaries of Sialidae and Raphidioptera. Both taxa belong to the Neuropterida. As in all telotrophic ovaries, all nurse cells are combined in an anterior chamber, the tropharium. The tropharium houses two subsets of germ cells: numerous nurse cell nuclei are combined in a central syncytium without any cell membranes in between, surrounded by a monolayer of single-germ cells, the tapetum cells. Each tapetum cell is connected to the central syncytium via an intercellular bridge. Tapetum cells of the posterior zone, which sufficiently contact prefollicular cells, are able to grow into the vitellarium and develop as oocytes. During previtellogenic and early vitellogenic growth, oocytes remain connected with the central syncytium of the tropharium via their anterior elongations, the nutritive cords. The morphological data are discussed in the light of those derived from ovaries of other Coleoptera and from the proposed sister group, the Neuropterida. The data strongly support a sister group relationship between Coleoptera and Neuropterida. Furthermore, several switches between polytrophic and telotrophic ovaries must have occurred during the radiation of ancient insect taxa.