Follicular epithelium, theca and egg envelope formation in Serrasalmus spilopleura (Teleostei, Characiformes, Characidae)

The follicular epithelium and theca of oocytes in Serrasalmus spilopleura differentiates during the initial primary growth phase. The follicular cells are squamous and the thecal cells are disposed in two layers. During the secondary growth phase, follicular cells become cuboidal, acquire characteristics typical of protein- or glycoprotein-producing cells, and show dilated intercellular spaces. Formation of the egg envelope in S. spilopleura begins in the previtellogenic oocytes as a layer of amorphous electron-dense material is laid down on the oolemma. During vitellogenesis, another layer of electron-dense material appears beneath the first layer. Also during this phase, a layer of amorphous, less electron-dense material is formed adjacent to the follicular epithelium. The secondary egg envelope appears at the postvitellogenic phase and is composed of a filamentous and undulant material. The morphology of the egg envelopes in S. spilopleura reflects not only its oviparous nature but also the fact that its eggs are adhesive.     

Corpus luteum formation and ovulation in the butterfly Calpodes

The first corpus luteum of each ovariole is formed initially from the epithelial plug of cells which underly the first leading oocyte through autolysis, characterized by increased acid phosphatase activity, autophagy and lipophilic material. Later autolysis spreads progressively to apposing follicular cells, degeneration moving upwards until the leading oocyte is in the ovariolar duct, resulting in ovulation. All subsequent corpora lutea are compound structures, comprising the degenerating follicle of the newly shed eggs as well as remnants of previous corpora lutea. The interfollicular bridge cells appear to envelope the mature follicle during corpus luteum formation, thereby giving some support to the ovariole as a follicle disintegrates und the egg is shed.     

The ovaries and changes in their structural components at the end of vitellogenesis and during vitelline membrane formation in the butterfly, Calpodes

The eight ovarioles of Calpodes ethlius are meroistic and polytrophic with seven nurse cells per follicle. The follicles consisting of oocyte, nurse cells and surrounding follicular cells are connected by interfollicular bridges, whose cells are characterized by bundles of microtubules which appear, with some fine filaments, to terminate on or near to the plasma membrane at hemidesmosomes. The ovariolar sheath consists of tightly knit circular and longitudinal muscles which are heavily tracheated. The ovariolar duct consists of more loosely arranged circular and longitudinal muscles and an inner epithelial layer, also tracheated. The vitelline membrane appears to be secreted largely by the oocyte first as an electron-lucent layer which becomes gradually more electron-dense, probably as a result of addition of material from the follicular cells. Overlapping plate-like structures on the outer surface of the fully formed vitelline membrane may provide waterproofing.     

Morphological changes in the oocyte and its surrounding vestments during in vivo fertilization in the dasyurid marsupial Sminthopsis crassicaudata

This light and transmission electron microscopical study shows that the first polar body is given off before ovulation and that part of its cell membrane and that of the surrounding oocyte have long microvilli at the time of its ejection. Several layers of cumulus cells initially surround the secondary oocyte and first polar body, but the ovulated oocytes in the oviducts in the process of being fertilized do not have cumulus cells around them. Partly expelled second polar bodies occur in the oviduct; they are elongated structures that lack organelles and have electron-dense nuclei. A small fertilization cone appears to form around the sperm tail at the time of sperm entry into the egg and an incorporation cone develops around the sperm head in the egg cytoplasm. In three fertilized eggs a small hole was seen in the zona, which was presumably formed by the spermatozoon during penetration. Cortical granules, present in ovarian oocytes, are not seen in fertilized tubal or uterine eggs; release of their contents probably reduces the chances of polyspermy, although at least one polyspermic fertilized egg was seen and several other fertilized eggs had spermatozoa within the zona pellucida. In the zygote the pronuclei come to lie close together, but there was no evidence of fusion. A "yolk mass," which becomes eccentric before ovulation, is extruded by the time the two-cell embryos are formed, but many vacuoles remain in the non-yolky pole of the egg. A shell membrane of variable thickness is present around all uterine eggs but its origin remains undetermined.     

Structure and morphogenesis of the eggshell and micropylar apparatus in the olive fly, Dacus oleae (Diptera: Tephritidae)

The egg of the olive fly, Dacus oleae (Diptera, Tephritidae), is laid inside olives and the larva eventually destroys the fruit. The oocyte is surrounded by several distinct layers which are produced during choriogenesis. The chorion covering the main body of the egg outside of the vitelline membrane includes a "wax" layer, an innermost chorionic layer, an endochorion consisting of inner and outer layers separated by pillars and cavities similar to their counterparts in Drosophila melanogaster, as well as inner and outer exochorionic layers. The anterior pole is shaped like an inverted cup, which is chiefly hollow around its base and has very large openings communicating with the environment. Holes through the surface of the endochorion result from deposition of endochorionic substance around follicular cell microvilli. An opening at the apex of the cup provides an entrance for sperm entering the micropylar canal, which traverses the endochorion and continues into a "pocket" in a thickened vitelline protrusion. The micropylar canal is formed by deposition of endochorion and vitelline membrane around an elongated pair of follicular cell extensions. These extensions later degenerate and leave an empty canal about 5 microns in diameter and the narrower pocket about 1 micron in diameter. Respiration is thought to be facilitated by openings at the base of the anterior pole as well as by openings through the "plastron" around the main body of the shell.     

Ovarian follicle ultrastructure in the teleost Synbranchus marmoratus (Bloch, 1795), with special reference to the vitelline envelope development

Synbranchus marmoratus is a protogynous diandric teleost fish widely distributed throughout South America. The aim of this work was to study the ultrastructure of the vitelline envelope and the relationship among oocyte and their follicular cells during oogenesis. During perinucleolar stage, the oocyte and the follicular cells form microvillar processes that project into the perivitelline space. The oocyte secretes a dense and amorphous material, which appears as the first evidence of the vitelline envelope (VE) development. The VE passes from a double to a multilayered structure during oocyte growth. In mature oocytes, the VE reach a mean thickness of 11 microm, having up to 30 layers. Oocyte microvilli are thinner than the follicular ones and were seen in contact with the follicular plasmalema, however we could not find any contact between the follicular microvilli and the oolemma. Before ovulation, microvillar processes retract and the pore canals seem to collapse. An outer electron dense layer occludes the superficial pore and forms a continuous layer. No jelly or adhesive coatings were seen at least in ovulated eggs sampled from ovarian lumen. Follicular cell and oocyte cytological characteristics do not differ from those described in other teleosts species.     

Oogenesis in the viviparous matrotrophic lizard Mabuya brachypoda

Oogenesis in the lizard Mabuya brachypoda is seasonal, with oogenesis initiated during May-June and ovulation occurring during July-August. This species ovulates an egg that is microlecithal, having very small yolk stores. The preovulatory oocyte attains a maximum diameter of 0.9-1.3 mm. Two elongated germinal beds, formed by germinal epithelia containing oogonia, early oocytes, and somatic cells, are found on the dorsal surface of each ovary. Although microlecithal eggs are ovulated in this species, oogenesis is characterized by both previtellogenic and vitellogenic stages. During early previtellogenesis, the nucleus of the oocyte contains lampbrush chromosomes, whereas the ooplasm stains lightly with a perinuclear yolk nucleus. During late previtellogenesis the ooplasm displays basophilic staining with fine granular material composed of irregularly distributed bundles of thin fibers. A well-defined zona pellucida is also observed. The granulosa, initially composed of a single layer of squamous cells during early previtellogenesis, becomes multilayered and polymorphic. As with other squamate reptiles, the granulosa at this stage is formed by three cell types: small, intermediate, and large or pyriform cells. As vitellogenesis progresses the oocyte displays abundant vacuoles and small, but scarce, yolk platelets at the periphery of the oocyte. The zona pellucida attains its maximum thickness during late oogenesis, a period when the granulosa is again reduced to a single layer of squamous cells. The vitellogenic process observed in M. brachypoda corresponds with the earliest vitellogenic stages seen in other viviparous lizard species with larger oocytes. The various species of the genus Mabuya provided us with important models to understand a major transition in the evolution of viviparity, the development of a microlecithal egg.     

Cell-to-cell communication and ovulation. A study of the cumulus-oocyte complex

Cell-to-cell communication was characterized in cumulus-oocyte complexes from rat ovarian follicles before and after ovulation. Numerous, small gap junctional contacts were present between cumulus cells and oocytes before ovulation. The gap junction are formed on the oocyte surface by cumulus cell processes that transverse the zona pellucida and contact the oolemma. The entire cumulus mass was also connected by gap junctions via cumulus-cumulus interactions. In the hours preceding ovulation, the frequency of gap junctional contacts between cumulus cells and the oocyte was reduced, and the cumulus was disorganized. Electrophysiological measurements indicated that bidirectional ionic coupling was present between the cumulus and oocyte before ovulation. In addition, iontophoretically injected fluorescein dye was tranferred between the oocyte and cumulus cells. Examination of the extent of ionic coupling in cumulus-oocyte specimens before and after ovulation revealed that ionic coupling between the cumulus and oocyte progressively decreased as the time of ovulation approached. In postovulatory specimens, no coupling was detected. Although some proteolytic mechanism may be involved in the disintegration of the cumulus-oocyte complex, neither the cumulus cells nor the oocyte produced detectable levels of plasminogen activator, a protease which is synthesized by membrana granulosa cells. In summary, cell communication is a characterisitc feature of the cumulus-oocyte complex, and this communication is terminated near the time of ovulation. This temporal pattern of the termination of communication between the cumulus and the oocyte may indicate that communication provides a mechanism for regulating the maturation of the oocyte during follicular development before ovulation.     

Oogenesis in a Flesh Fly, Sarcophaga ruficornis

Maturation of the oocyte in the polytrophic ovariole of the flesh fly, Sarcophaga ruficornis was subdivided into 12 growing stages. The nurse cells increasing in volume during initial stages 1–6, shrink sharply in size from stage 7 onwards and finally disintegrate in stage 12. The oocyte continues to increase in volume till it reaches 1.2×107⁷μ³ in stage 12 before ovulation. The onset of vitellogenesis is marked by the appearance of protein-containing granules in the peripheral ooplasm in stage 7. The follicular epithelial cells around the oocyte exhibit changes in their shape in different stages and perhaps facilitate as well as stop the transport of yolk precursors to the ooplasm. Finally, after the formation of egg membranes around the yolk-filled oocyte, the follicular epithelium is sloughed off in stage 12.     

The origin and structure of the zona pellucida in the ovarian eggs of teleosts

Summary An exhaustive study of the egg membranes of the oocytes of Trichiurus savala and Triacanthus brevirostris was made with particular reference to the origin and structure of the zona pellucida. The zona pellucida makes its first appearance as a deeply stained narrow zone of fine granular or amorphous homogeneous material around the oocytes, between the follicular epithelium and the vitelline membrane. This homogeneous substance is the product of the follicle cells. Very soon, radial striations in the zona pellucida get formed out of the homogeneous substance of the zona itself. The peripheral cytoplasm of the oocyte, by this time, also gets differentiated into a fibrillar layer. The follicle cells as well as the ooplasmic fibrillar layer send out a number of protoplasmic fibres towards each other into the canalicular passages of the radially striated zona. These fibres meet in the middle of the zona, differentiating it into two concentric zones, an outer—zona radiata externa and an inner-zona radiata interna. The former is purely follicular in origin, whereas the latter is partly follicular and partly ooplasmic in nature.     

Influence of hormones on the inhibitory activity of oocyte maturation present in conditioned media of porcine granulosa cells

This study examines the influence of follicular maturation as well as the role of various hormones upon the secretion of an oocyte maturation inhibitor (OMI) from porcine granulosa cells incubated in vitro. The results demonstrate that the OMI substance, secreted into the media by granulosa cells, is present in a low molecular-weight fraction (< 10,000 daltons) similar to that found in follicular fluid of porcine antral follicles. Also, as follicular development progresses, the granulosa cells lose their ability to secrete OMI. More importantly, hormones appear to regulate OMI secretion: FSH stimulates OMI secretion and androgens inhibit OMI secretion. These data provide evidence for the proposal of the following hypothesis concerning hormonal regulation of oocyte, meiosis by OMI in the porcine follicle: Whether the oocyte resumes meiosis, either during atresia or ovulation, is dependent upon the proper milieu of gonadotropins, cyclic-AMP, and steroids within the microenvironment of the follicular compartment. The cellular interactions of these hormones, particularly FSH and androgens, control the amount of OMI (and possibly other intrafollicular factors) secreted in the follicle, which may be involved in either maintaining the immature state or permitting meiotic maturation.     

Egg structure of Zorotypus caudelli Karny (Insecta, Zoraptera, Zorotypidae)

The structural features of eggs of Zorotypus caudelli Karny are described in detail. The egg is elliptic with long and short diameters of 0.6 and 0.3 mm respectively, and creamy white. The egg shows a honeycomb pattern on its surface, without any specialized structures for hatching such as an operculum or a hatching line. The fringe formed by a fibrillar substance secreted after the completion of the chorion encircles the lateral surface. The egg layer is composed of an exochorion, an endochorion, and a vitelline envelope. The exochorion and endochorion are electron-dense and homogeneous in structure. The exochorion shows a perforation of numerous branching aeropyles. The exo- and endochorion are connected by numerous small columnar structures derived from the latter. The vitelline envelope is very thin and more electron-dense than the chorion. A pair of micropyles is present at the equator on the dorsal side of the egg. Originating at the micropyle, the micropylar canal runs through the chorion obliquely. The structural features of the eggs of Zoraptera were compared with those of other polyneopteran and paraneopteran orders.     

The development of porcine zona pellucida using monoclonal antibodies: II. Electron microscopy

Following our previous study on the immunohistochemistry of porcine zonae pellucidae (ZP), we undertook the present study to localize the components of the ZP with immunoelectron microscopy, using three types of anti-porcine-ZP monoclonal antibodies (Mabs), named STA-1, STA-2, and STA-3. Some organelles of the oocyte were seen to react with STA-2 and STA-3 prior to ZP formation. As soon as a follicle began to mature, STA-2 and STA-3 reacted with the perinuclear space and the endoplasmic reticular membrane of the oocyte. The follicle first reacted with STA-1 at the secondary follicle stage. At this stage, the positive reaction involved the follicular cell layer as well as the oocyte and ZP. Positive reaction was scattered within and limited to the interfollicular cell space and was never found in the cytoplasm of follicular cells. At the antral follicle stage, the oocyte was surrounded by a thick, electron-dense ZP. A strong reaction was observed in the outer layer, but no significant reaction occurred in the inner layer. The convex and ragged outer margin of the ZP was characterized by the strongest reaction.     

Oogenesis: From Oogonia to Ovulation in the Flagfish,Jordanella floridae Goode and Bean, 1879 (Teleostei: Cyprinodontidae)

We provide histological details of the development of oocytes in the cyprinodontid flagfish, Jordanella floridae. There are six stages of oogenesis: Oogonial proliferation, chromatin nucleolus, primary growth (previtellogenesis [PG]), secondary growth (vitellogenesis), oocyte maturation and ovulation. The ovarian lamellae are lined by a germinal epithelium composed of epithelial cells and scattered oogonia. During primary growth, the development of cortical alveoli and oil droplets, are initiated simultaneously. During secondary growth, yolk globules coalesce into a fluid mass. The full‐grown oocyte contains a large globule of fluid yolk. The germinal vesicle is at the animal pole, and the cortical alveoli and oil droplets are located at the periphery. The disposition of oil droplets at the vegetal pole of the germinal vesicle during late secondary growth stage is a unique characteristic. The follicular cell layer is composed initially of a single layer of squamous cells during early PG which become columnar during early vitellogenesis. During primary and secondary growth stages, filaments develop among the follicular cells and also around the micropyle. The filaments are seen extending from the zona pellucida after ovulation. During ovulation, a space is evident between the oocyte and the zona pellucida. Asynchronous spawning activity is confirmed by the observation that, after ovulation, the ovarian lamellae contain follicles in both primary and secondary growth stages; in contrast, when the seasonal activity of oogenesis and spawning ends, after ovulation, the ovarian lamellae contain only follicles in the primary growth stage. J. Morphol. 277:1339–1354, 2016. © 2016 Wiley Periodicals, Inc.     

Ultrastructural studies on the nematode Xiphinema diversicaudatum: Egg shell formation

The ultrastructure of the formation of the egg shell in the longidorid nematode Xiphinema diversicaudatum is described. Upon fertilization a vitelline membrane, which constitutes the vitelline layer of the egg shell, is formed. The chitinous layer is secreted in the perivitelline space, between the vitelline layer and the egg cell membrane. On completion of the chitinous layer, the material of the lipid layer is extruded from the egg cytoplasm to the outer surface, through finger-like projections. Both chitinous and lipid layers are secreted by granules in the egg cytoplasm that disappear as the layers are completed. Chitinous and lipid layers are formed during the passage of the egg through the oviduct. The vitelline layer is enriched with secretions produced by the oviduct cells and then by phospholipids secreted by the cells of the pars dilatata oviductus. The inner uterine layer is also formed by deposition of secretory products apposed on the egg shell in the distal uterine region and Z-differentiation. In the proximal part of the uterus, the egg has a discontinuous electron-dense layer, the external uterine layer. Tangential sections between chitinous and uterine layers revealed the presence of holes, possibly egg pores, delimited by the two uterine layers.     

Egg Envelope Cytodifferentiation in the Colonial Ascidian Botryllus schlosseri (Tunicata)

Abstract The formation and cytodifferentiation of egg envelopes were studied at the ultrastructural level in blastozooids of Botryllus schlosseri. The process was divided into five recognized stages of oogenesis. First, the small young oocytes (stage 1) are contacted by scattered cells (primary follicle cells—PFC) which adhere to the oolemma at several junctional spots. PFC extend all around the growing oocyte, acquire polarity, and form a layer covered externally by a thin basal membrane (stage 2). At stage 3 isolated cells are recognizable between the PFC layer and oocyte. They never form junctions with the oocyte and represent prospective inner follicle cells (IFC) and test cells (TC), the latter being progressively received in superficial depressions in the oocyte. The layer of PFC, which maintains junctions with the oolemma, represents prospective outer follicle cells (OFC). PFC are considered to be the source of the three cellular envelopes because a contribution from mesenchymatous elements was not observed. At the beginning of vitellogenesis (stage 4), the vitelline coat (VC) becomes recognizable as a loose net covering the oocyte and TC. It is crossed by the oocyte microvilli and OFC projections which meet and form numerous small junctional plaques, some of them resembling gap junctions. IFC, VC and TC show marked signs of differentiation with approaching ovulation. OFC differentiate completely before ovulation (stage 5) and are engaged in intense synthesis of proteins which may be transferred and taken by endocytosis into the oocyte for yolk formation. Experiments with injected horseradish peroxidase also revealed that proteins present in the blood may reach the oocyte via the intercellular pathway, overcoming OFC and IFC. The possible roles of all the egg envelopes are discussed.     

Early oogenesis in the ascidian Ciona savignyi

Summary

Morphology of the germinal epithelium and the early follicular oocyte in the ascidian Ciona savignyi as examined by electron microscopy. The oogenetic part of the germinal epithelium contains oocytes at two different stages and the dark and clear cells. The smaller oocyte contains synaptonemal complexes. The larger oocyte in the initial phase of growth has a conspicuous nucleolus, electron-dense materials and some mitochondria close to the nuclear envelope. The nucleus of the larger oocyte is round and has the smooth contour. The dark cell contains a relatively large nucleus and is sometimes connected to each other by an intercellular bridge. Therefore, the dark cell, which has been suggested to be the progenitor cell of two kinds of accessory cells, may be also the oogonium. The early follicular oocyte just after migration from the germinal epithelium retains most of cytological features similar to those of the larger oocyte. However, the nuclear contour of the early follicular oocyte is uneven. This difference in the nuclear contour probably indicates that such a follicular oocyte is in the second phase of growth.     

The development of the hexagonally structured egg envelope of the C-O sole (Pleuronichthys coenosus)

The surface of a mature, pelagic C-O sole egg is composed of polygonal chambers having four to eight sides, most of which are hexagonally shaped. This honeycomb pattern initially appears on primary oocytes as a thin layer of compact, electron-dense material. Discrete thickenings begin to develop on the envelope of perinuclear stage oocytes. The thickenings lengthen and thin to form the hexagonal walls of the envelope in oocytes undergoing yolk vesicle formation. The walls of each hexagonal chamber occur in an area corresponding to the lateral margins of the adjacent follicle cell, suggesting that the hexagonal walls are produced by the follicle cells. The hexagonal layer is nearly complete at the beginning of vitellogenesis, and as vitellogenesis continues, a striated envelope layer composed of fibrillar lamellae develops between the oocyte and the hexagonal layer. The striated layer appears to be secreted by the oocyte. After vitellogenesis is completed, oocytes are ovulated and double in size during a period of maturation. Concurrently, the striated primary envelope stretches and thins into eight to nine horizontal lamellae. On the mature egg surface, the polygonal chambers are about 24–31 μm in diameter. Within each chamber there is a subpattern of polygonal areas; each polygon is 1.5–2.0 μm in diameter, and circumscribes a pore canal opening. This exceptional envelope may furnish the egg with some degree of protection, resiliency, and buoyancy, but its specific functions are not known.     

The eggshell of the almond wasp Eurytoma amygdali (Hymenoptera, Eurytomidae) - 2. The micropylar appendage

Micropylar apparatuses in insects are specialized regions of the eggshell through which sperm enters the oocyte. This work is an ultrastructural study and deals with the structure and morphogenesis of the micropylar appendage in the hymenopteran Eurytoma amygdali. The micropylar appendage is a 130 mum long cylindrical protrusion located at the posterior pole of the egg, unlike other insects i.e. Diptera. in which the micropylar apparatus is located at the anterior pole. In mature eggs there is a 0.4 mum wide pore (micropyle) at the tip of the appendage leading to a 6 mum wide micropylar canal. The canal contains an electron-lucent substance, it travels along the whole appendage and finally reaches the vitelline membrane of the oocyte. The vitelline membrane is covered by a wax layer and an electron-lucent layer, whereas the chorion surrounding the canal consists of a granular layer (fine and rough) and a columnar layer. The morphogenesis of the appendage starts in immature follicles: four central cells located at the posterior tip of the oocyte near the vitelline membrane, differing morphologically from the adjacent follicle cells. These central cells degenerate during early chorionic stages, thus assisting in the formation of the micropylar canal. The adjacent, peripherally located cells secrete the electron-lucent substance which fills the canal and at the same time, the fine granular layer is formed starting from the base towards the tip of the appendage. The secretion persists at late chorionic stages and results in the formation of the chorion around the micropylar canal. The extremely long (compared to other insects) micropylar appendage seems to facilitate the egg passage through the very thin and long ovipositor. The structure and morphogenesis of this appendage differs significantly from the micropylar apparatuses studied so far in other insects i.e. Diptera, and may reflect adaptational and evolutionary relationships.     

Evidence for Follicle Wall Involvement in Ovulation and Progesterone Production by Frog (Rana pipiens) Follicles in vitro

Involvement of different cellular investments of the amphibian ovarian follicle wall in the ovulatory process, progesterone production, and oocyte maturation was investigated. Following microdissection, to selectively remove one or more layers (surface epithelium, theca, follicle cells) of the follicle wall, dissected and undirected ovarian follicles were treated with frog pituitary homogenate (FPH) or progesterone. Intact follicles ovulated in response to pituitary homogenate and this was associated with contractions of the follicle wall. Ovulation and follicular contractions were not observed following removal of the surface epithelium without removing the thecal layer. Oocyte maturation occured in response to FPH following removal of the surface epithelium alone or together with the theca, but not in the absence of the follicle cells. Intact follicles were most responsive to FPH with respect to progesterone production, and removal of all somatic cells from oocytes obliterated FPH stimulated progesterone production. Oocytes, regardless of wether any or all follicular wall layers were removed, matured but did not ovulate following exposure to progesterone. The results suggest that the surface epithelium, but not the theca, is required for FPH-induced extrusion (ovulation) of the oocyte from ovarian follicle wall. Additionally, the somatic tissue rather than the oocyte appears to be the cells producing progesterone following FPH treatment. The results indicate that separate cellular layers (individually and/or as a result of interactions) of the follicle wall carry out different functions during follicular differentiation and mediation of ovulation. Data provide functional evidence for a role of the surface epithelium in controlling the process of ovulation and follicular contraction.