Structure of ovaries and formation of egg envelopes in the stonefly,Leuctra autumnalis Aubert, 1948 (Plecoptera: Leuctridae). Ultrastructural studies

The investigation of ovaries and the formation of egg envelopes of the stonefly Leuctra autumnalis was carried out with light and transmission electron microscopes. The ovary of the studied species is paired and consists of several dozen panoistic ovarioles opening individually to the oviduct. The process of egg capsule formation already begins in previtellogenesis. At this time the follicular cells secrete precursors of the vitelline envelope. Analysis of the presented data suggests that the oocyte itself also takes part in the formation of the vitelline envelope during late vitellogenesis. Simultaneously, the follicular cells produce precursors of further layers of the egg capsule, i.e. two-layered chorion and extrachorion, consisting of two gelatinous layers and a flocculent one. The completely developed capsule contains channels, probably micropylar ones.     

Formation and structure of egg capsules in scale insects (Hemiptera, Coccinea) I. Ortheziidae

The paired ovaries of the investigated species are composed of 20-30 ovarioles of a telotrophic-meroistic type. Each ovariole is subdivided into an apical tropharium (=trophic chamber) and a vitellarium that contains a single developing oocyte. This oocyte is surrounded by a mono-layered follicular epithelium that is responsible for synthesis of precursors of egg envelopes. In Orthezia, synthesis and secretion of precursors of egg envelopes (=choriogenesis) and accumulation of reserve substances in the oocyte cytoplasm (=vitellogenesis) start at the same time. The egg capsule is composed of two envelopes: an internal, thick vitelline envelope and an external, very thin chorion. The egg surface is covered with numerous, irregularly arranged waxy filaments of spiral shape. Eggs are devoid of the micropylar, aeropylar and hydropylar openings.     

Ultrastructural studies of the formation of the egg capsule in the hermaphroditic species, Isohypsibius granulifer granulifer Thulin, 1928 (Eutardigrada: Hypsibiidae)

The egg capsule of Isohypsibius granulifer granulifer Thulin 1928 (Eutardigrada: Hypsibiidae) is composed of two shells: the thin vitelline envelope and the multilayered chorion. The process of the formation of the egg shell begins in middle vitellogenesis. The I. g. granulifer vitelline envelope is of the primary type (secreted by the oocyte), but the chorion should be regarded as a mixed type: primary (secreted by the oocyte), and secondary (produced by the cells of gonad wall). During early choriogenesis, the parts of the chorion are produced and then connected into a permanent layer. The completely developed chorion consists of three layers: (1) the inner, medium electron dense layer; (2) the middle labyrinthine layer; (3) the outer, medium electron dense layer. After the formation of the chorion, a vitelline envelope is secreted by the oocyte.     

Eggshell fine structure of Bradysia aprica (Winnertz) (Diptera : Sciaridae)

The eggshell fine structure of the dark-winged fungus-gnat Bradysia aprica (Winnertz) (Diptera : Sciaridae) was investigated by scanning and transmission electron microscopy. At the anterior pole of the ovoid egg is a single micropyle, centrally located in a well-defined micropylar area. The latter is covered by many long drumstick-like chorionic processes that are longer and more numerous than those of the rest of the egg surface. Cross-sections of the eggshell show 3 concentric envelopes: the vitelline envelope, wax layer and chorion. The chorion consists of 3 components with different morphological features: the inner, intermediate and outer chorion. The latter 2 layers, involved in the organization of the drumstick-like processes, have homogeneous features, whereas the former is crystalline and resembles the innermost chorionic layer of other Diptera.     

Previtellogenic and vitellogenic oocytes in ovarian follicles of cultured siberian sturgeon Acipenser baerii (Chondrostei,Acipenseriformes)

Previtellogenic and vitellogenic oocytes in ovarian follicles from cultured Siberian sturgeon Acipenser baerii were examined. In previtellogenic oocytes, granular and homogeneous zones in the cytoplasm (the ooplasm) are distinguished. Material of nuclear origin, rough endoplasmic reticulum, Golgi complexes, complexes of mitochondria with cement and round bodies are numerous in the granular ooplasm. In vitellogenic oocytes, the ooplasm comprises three zones: perinuclear area, endoplasm and periplasm. The endoplasm contains yolk platelets, lipid droplets, and aggregations of mitochondria and granules immersed in amorphous material. In the nucleoplasm, lampbrush chromosomes, nucleoli, and two types of nuclear bodies are present. The first type of nuclear bodies is initially composed of fibrillar threads only. Their ultrastructure subsequently changes and they contain threads and medium electron dense material. The second type of nuclear bodies is only composed of electron dense particles. All nuclear bodies impregnate with silver, stain with propidium iodide, and are DAPI‐negative. Their possible role is discussed. All oocytes are surrounded by follicular cells and a basal lamina which is covered by thecal cells. Egg envelopes are not present in previtellogenic oocytes. In vitellogenic oocytes, the plasma membrane (the oolemma) is covered by three envelopes: vitelline envelope, chorion, and extrachorion. Vitelline envelope comprises four sublayers: filamentous layer, trabecular layer 2 (t2), homogeneous layer, and trabecular layer 1 (t1). In the chorion, porous layer 1 and porous layer 2 are distinguished in most voluminous examined oocytes. Three micropylar cells that are necessary for the formation of micropyles are present between follicular cells at the animal hemisphere. J. Morphol. 278:50–61, 2017. ©© 2016 Wiley Periodicals,Inc.     

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.     

Egg-shells in mites

Summary This communication presents results of studies on the formation and structure of the vitelline envelopes in three species of mites: Euryparasitus emarginatus (Gamasida), Erythraeus phalangoides (Actinedida), and Hafenrefferia gilvipes (Oribatida). In E. emarginatus and E. phalangoides, in which the oocytes are not covered with follicular cells, the material of the vitelline envelope appears first in vesicles under the surface of the oocytes prior to secretion by exocytosis. The formed vitelline envelope is built of a homogeneous material which is perforated by numerous channels containing oocyte microvilli. Later, as the microvilli are retracted, the channels disappear. In both of these species the formed vitelline envelope is incomplete and the micropylar orifice occurs as a transitional structure.In H. gilvipes follicular cells encircling the oocyte contain granules filled with material that is subsequently secreted into the perivitelline space forming the vitelline envelope on the oocyte surface. The inner layer of the vitelline envelope is granular, whereas the outer part is more homogeneous. Both lack channels containing microvilli and micropyle.     

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.     

Egg envelopes in diplopods, a comparative ultrastructural study

By means of electron microscopy two types of egg envelope have been described in representatives of two diplopod subclasses, the Chilognatha and the Pselaphognatha. The vitelline envelope appears on the oocyte surface in early previtellogenesis and persists till ovulation. In its thin and filamentous structure it resembles basement membranes. During vitellogenesis electron dense material is deposited on filamentous scaffolding which fills the space between the oolemma and the vitelline envelope. As a result, the thick and spongy or filamentous chorion is formed. In the present study it has been shown that regardless of the type of oogenesis (solitary-the Chilognatha, or follicular-the Pselaphognatha) both envelopes in diplopods are produced by the oocyte itself, and although completely different in structure and time of appearance, they must be both considered as primary.     

The vitelline envelope,chorion, and micropyle of Fundulus heteroclitus eggs

The architecture and transformation of the vitelline envelope of the developing oocyte into the chorion of the mature egg of Fundulus heteroclitus have been examined by scanning and transmission electron microscopy. The mature vitelline envelope is structurally complex and consists of about nine strata. The envelope is penetrated by pore canals that contain microvilli arising from the oocyte and macrovilli from follicle cells. During the envelope's transformation into the chorion, the pore canals are lost and the envelope becomes more fibrous and compact and its stratified nature less apparent. The micropyle, of pore, through which the sperm gains access to the enclosed egg is located at the bottom of a small funnel-shaped depression in the envelope. Internally, the micropyle opens on the apex of a cone-like elevation of the chorion. During the development of the envelope, structured chorionic fibrils, the components of which are presumed to be synthesized by the follicle cells, become attached to its surface. These chorionic fibrils are though to aid in the attachment of the egg to the substratum and perhaps to help prevent water loss during low tides when the egg may be exposed.     

Micro analysis of the egg shell of Adela metallica (Poda) (Lepidoptera : Adelidae) by energy-filtering transmission electron microscopy (EFTEM)

The egg shell of the incurvarioid moth Adela metallica (Lepidoptera : Adelidae) was studied by conventional (cTEM) and energy-filtering transmission electron microscopy (EFTEM). The shell of the laid egg consists of 3 envelopes. The vitelline envelope is 0.1–0.2μm thick and homogeneous, thus exhibiting the non-exoporian character state. The single-layered chorion, which is covered by a fibrogranular mucous layer, is 0.5–0.9μm thick and homogeneous, thus exhibiting the non-ditrysian character state. The chorion is highly electron-lucent. Neither cTEM nor EFTEM revealed any sub-structural details. However, electron spectroscopic imaging (ESI) and electron energy-loss spectroscopy (EELS), revealing the elemental composition of the egg shell, indicate that the chorion and vitelline envelope are proteinaceous and hence, similar to the egg shells of other lepidopteran species. The presence of high sulphur signals associated with the vitelline envelope and the thin basal lamella of the chorion indicates that these components may be stabilized via sulphur-bridges.     

Formation and ultrastructure of the micropylar apparatus in Bombyx mori ovarian follicles

The formation of the micropylar apparatus during oogenesis in the silkworm, Bombyx mori, has been studied using light and transmission electron microscopy. The micropylar apparatus is formed by three types of cells: the micropylar channel-forming cells (MCFCs), the micropylar orifice-forming cells (MOFCs), and the micropylar rosette-forming cells (MRFCs). During the formation of the vitelline membrane and the chorion, each of the MCFCs extends a cytoplasmic projection serving as the mold of a micropylar-channel into the egg envelopes. The detachment and collapse of the projections takes place at the end of choriogenesis. The micropylar channels possess a common external orifice on the chorion and several internal orifices within the vitelline membrane. The MOFCs interact closely with the MCFCs and contribute to the formation of the external micropylar orifice. A petal-like rosette surrounding the orifice is imprinted on the outer chorionic surface by the MRFCs which enclose a group of the MCFCs and MOFCs.     

Ultrastructure and resistance to water loss in eggs of Acanthoscelides obtectus Say (Coleoptera: Bruchidae)

The mature oöcyte of Acanthoscelides obtectus is surrounded by three envelopes: an external layer, a chorion and a vitelline membrane. The external layer is secreted by the walls of the lateral oviducts. The chorion and vitelline membrane are secreted by the follicular cells. The vitelline membrane becomes very compact during the hour following fertilization and laying. The chorion is composed of three layers, one of which has a paracrystalline ultrastructure.Mature, unfertilized, chorion-containing oöcytes, whose vitelline membranes are loose, dehydrate rapidly in a dry atmosphere after laying or after removal from the lateral oviducts. Fertilized eggs are quite resistant to desiccation: after 12 days at 25°C and 5% relative humidity, viable larvae are obtained.The compact vitelline membrane is the most effective protection against dehydration. The chorion and the external layer are much less effective in preventing water loss from the egg.The retention of eggs in the lateral oviducts does not seem to lead to any modification of the structure of their envelopes.     

Fine structure of the eggshell of Ommatissus binotatus Fieber (Homoptera,Auchenorrhyncha, Tropiduchidae)

The external morphology and fine structure of the eggshell of Ommatissus binotatus Fieber (Homoptera : Tropiduchidae) was investigated by light, scanning and transmission electron microscopy. The egg surface has 2 main regions: a specialized area and an unspecialized egg capsule. The specialized area is characterized by a large respiratory plate containing the operculum and a short respiratory horn. The latter consists of an external hollow tube and an internal coneshaped projection hosting a micropylar canal. The eggshell has 4 layers: the vitelline envelope, a wax layer, the chorion and an outer mucous layer. The chorion has inner, intermediate and outer parts. The functions of the different parts of the eggshell are discussed. Characters useful to define the eggs and the oviposition habit in the family Tropiduchidae were provided. The size and morphology of the egg, plate, respiratory horn and operculum are suggested as useful characters for ootaxonomic analysis.     

Cytochemical studies of developmental stages during oogenesis in Culex pipiens molestus (Forsk?l). I. Lipids and carbohydrates

Lipids and carbohydrates were studied in the polytrophic ovaries of Culex pipiens molestus during oogenesis. The cytoplasm of both the oocyte and the nurse cells contains lipid structures at all stages of development--granules in the early stages and spheres in the later stages. Intranuclear lipid bodies can be demonstrated in the oocyte and in the nurse cells. After leaving the nucleus, lipids are deposited in the peripheral cytoplasm. From the third to the seventh adult phase, lipid granules are concentrated in the area of the nurse cell and oocyte junction, indicating that lipids originate in the nurse cells and are transported from these to the oocyte. The follicular epithelial cells provide the oocyte with lipid material for fatty yolk synthesis and formation of the egg envelopes. Lipids are distributed similarly to the Golgi apparatus, indicating that there is a relationship between this organelle and fat formation. In the early stages, the cytoplasm of the oocyte, the nurse cells and the follicular epithelium contains glycogen granules. In the later stages these cells also contain mucopolysaccharides. The mucopolysaccharide yolk spheres are enclosed in vacuoles, while the chorion is composed of acid mucopolysaccharides. The follicular epithelium and vitelline membrane are of a mucopolysaccharide nature. A topographical relationship exists between the Golgi apparatus and the glycogen granules, indicating that this organelle also plays a role in glycogen synthesis.     

Ultrastructural features and formation of the micropylar apparatus in the cherry fly Rhagoletis cerasi

The micropylar apparatus (MA) in Rhagoletis cerasi (Diptera, Tephritidae) is located at the anterior pole of the egg and consists of two parts: an outer chorion and an inner vitelline membrane. Sperm entry takes place through the micropylar canal, 2.0–2.5 μm in diameter, which penetrates the micropylar endochorion and terminates in the thick vitelline membrane, thus forming the “pocket.” The pore of the micropylar canal, i.e., the micropyle, is covered by the exochorionic tuft. The formation of the MA is accomplished by 40 micropylar cells during oogenesis. These cells secrete the successive eggshell layers: the vitelline membrane, the wax layer, the innermost chorionic layer, the micropylar endochorion, and the exochorion. Two among 40 micropylar cells differentiate and form two tightly connected projections. The latter contain a bundle of parallel microtubules and participate in the formation of the micropylar canal and the pocket. At the tip of the projections there are two thin extensions full of microfilaments. In late developmental stages the two projections and the extensions degenerate and leave the canal and the pocket behind. We also discuss the structural features of the MA in relation to its physiology among Diptera.     

The ultrastructure and function of follicle cells in Foucartia squamulata (Herbst) (Curculionidae)

Summary The follicle cells of Foucartia squamulata are involved in the formation of both vitelline membrane and chorion. Precursors for these egg coverings are synthesized by the rough endoplasmic reticulum and condensed within dictyosomes. The vitelline membrane and the chorion appear on the oocyte surface simultaneously, which is an unusual phenomenon for insects. The follicular epithelium has not been found to contribute to vitellogenesis in the species under study.     

Eggshell fine structure of three lepidopteran pests: Cydia pomonella (L.) (Tortricidae), Heliothis virescens (Fabr.), and Spodoptera littoralis (Boisd.) (Noctuidae)

The eggshells of 3 moths, Cydia pomonella (Tortricidae), Heliothis virescens, and Spodoptera littoralis (Noctuidae) were investigated by scanning (SEM) and transmission (TEM) electron microscopy. The surface of the noctuid eggs shows structural elements (micropylar rosette, ribs, cross-ribs, and aeropyles) and regional differentiation, all typical of Lepidoptera. The egg of C. pomonella shows a different regional morphology due to its watch-glass shape and its position, lying on the flank. The micropylar structures are on the lower egg face in contact with the substrate. For S. littoralis, the surface structure (sculpturing) of the egg is not species-specific, being indistinguishable from that of S. frugiperda (Salkeld, 1984).In all 3 moths, the eggshell fine structure is basically identical, as revealed by TEM. Both the vitelline envelope and the chorion consist of several distinct layers. The vitelline envelope, bi-layered and several μm thick, undergoes a marked structural change when embryogenesis begins. At the same time, Golgi vesicles bearing dense particles, appear in the periplasm of the egg cell in fertilized eggs of H. virescens and S. littoralis. The chorion of all 3 species consists of a basal layer (C-1), a cavity layer (C-2) supported by trabecles and opening to the exterior via aeropylar canals, and a lamellar layer (C-3), which probably consists of helicoidally arranged stacks of fibrils. In H. virescens and S. littoralis, an additional epicuticle-like layer (C-4) is present. Available data from the literature are summarized and a basic scheme of the radial eggshell fine structure of ditrysian Lepidoptera is proposed.     

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.     

Fine structure and morphogenesis of the micropylar apparatus in the medfly Ceratitis capitata (Wiedemann) (Diptera : Tephritidae)

The micropylar apparatus (MA) in Ceratitis capitata (Diptera : Tephritidae) is a cone-like protrusion, 18 μm long, at the anterior pole of the egg, and exhibits about 40 follicle cell imprints externally. It consists of chorionic and vitelline membrane parts. The first contains at least a 3 μm wide micropylar canal; the tip of the MA is covered by a “tuft” and includes the micropyle, i.e. the entrance of the micropylar canal. The canal leads to the vitelline membrane part, where it forms a pocket. The sperm enters the oocyte by passing through the micropyle-micropylar canal-pocket route.At least 40 follicle cells participate in the formation of the micropylar apparatus. Two of these form 2 projections, which are tightly connected, and serve as a template for the formation of the canal and the pocket. Throughout their length, both projections have microtubules in parallel arrangement. During oogenesis, the remaining micropylar cells secrete the successive eggshell layers, i.e. the vitelline membrane, the wax layer, the innermost chorionic layer, the endochorion, and the exochorion. Towards the end of oogenesis, the 2 projections degenerate, and the canal becomes available for sperm passage.