Vitellogenesis of the digenean Plagiorchis elegans (Rudolphi, 1802) (Plagiorchioidea,Plagiorchiidae)

The ultrastructural organization of vitellogenesis of Plagiorchis elegans (Rudolphi, 1802), experimentally obtained from the golden hamster Mesocricetus auratus (Linnaeus, 1758), is described using transmission electron microscopy. This study is the first ultrastructural study of vitellogenesis in a member of the superfamily Plagiorchioidea. The four stages usually observed during vitellogenesis are described: stage I, cytoplasm of the vitellocytes mainly filled with ribosomes and few mitochondria; stage II, beginning of the synthetic activity; stage III, active synthesis of the shell globule clusters; stage IV, vitellocytes are filled with shell globule clusters and contain several lipid droplets, and glycogen granules are grouped around clusters and droplets. Vitellogenesis in P. elegans is compared with that of other Digenea. The differences among P. elegans and previously studied digeneans include, but are not limited to the occurrence of dense coiled endoplasmic reticulum saccules and the concentration of glycogen in the mesenchyme, which may be considered as a fifth stage of maturation of the vitelline glands. This peculiarity was not observed in all trematodes, which clearly indicates differences in the vitellogenesis in various digenean lineages at different stages of maturation of their vitelline cells.     

First study of vitellogenesis of the broad fish tapeworm Diphyllobothrium latum (Cestoda,Diphyllobothriidea), a human parasite with extreme fecundity

In the present study, the process of vitellogenesis of one of the most prolific organisms, the broad tapeworm, Diphyllobothrium latum, the causative agent of human diphyllobothriosis, was studied for the first time using transmission electron microscopy. Cytochemical staining with periodic acid-thiosemicarbazide-silver proteinate for detection of glycogen was applied. Starting from the periphery toward the center of the vitelline follicle four stages of vitellocytes are differentiated: immature vitellocytes, early maturing vitellocytes, advanced maturing and mature vitellocytes. Differentiation into mature vitellocytes involves the formation of shell globule clusters containing shell globules, large amount of saturated lipid droplets and glycogen. A peculiar ultrastructural feature of D. latum vitellogenesis is the presence of lamellar bodies in the cytoplasm of mature vitellocytes. This feature is similar to that present in the closely related caryophyllideans and spathebothriideans. Despite the great similarity observed in the embryonic development of diphylobothriideans, caryophyllideans and spathebothriideans, and the fact that their vitellocytes share a feature not reported from other cestode groups, there are substantial differences in the morphology of vitelline clusters, types, amount and localization of their nutritive reserves.     

Ultrastructure of vitellogenesis and vitellocytes in the trypanorhynch cestode Aporhynchus menezesi, a parasite of the velvet belly lanternshark Etmopterus spinax

This is the first TEM examination of vitellogenesis in the cestode Aporhynchus menezesi, a parasite of the velvet belly lanternshark Etmopterus spinax and a member of a little-studied trypanorhynch family, the Aporhynchidae. The synthetic activity of vitellocytes plays two important functions in the developmental biology of cestodes: (1) their shell-globules serve in eggshell formation; and (2) their accumulated reserves of glycogen and lipids represent a food source for the developing embryo. In A. menezesi, vitelline follicles consist of cells at various stages of development, from peripheral, immature cells of the gonial type to mature cells towards the centre of the follicle. These stages are: (I) immature; (II) early differentiation; (III) advanced maturation; and (IV) mature. Gradual changes involved in this process occur within each stage. Vitellogenesis involves: (1) an increase in cell volume; (2) the development of a smooth endoplasmic reticulum and an accelerated formation and accumulation of both unsaturated and saturated lipid droplets, along with their continuous enlargement and fusion; (3) the formation of individual β-glycogen particles and their accumulation in the form of glycogen islands scattered among lipid droplets in the cytoplasm of maturing and mature vitellocytes; (4) the rapid accumulation of large, moderately saturated lipid droplets accompanied by dense accumulations of β-glycogen along with proteinaceous shell-globules or shell-globule clusters in the peripheral layer during the advanced stage of maturation; (5) the development of cisternae of granular endoplasmic reticulum that produce dense, proteinaceous shell-globules; (6) the development of Golgi complexes engaged in the packaging of this material; and (7) the progressive and continuous enlargement of shell-globules into very large clusters in the peripheral layer during the advanced stage of maturation. Vitellogenesis in A. menezesi, only to some extent, resembles that previously described for four other trypanorhynchs. It differs in: (i) the reversed order of secretory activities in the differentiating vitellocytes, namely the accumulation of large lipid droplets accompanied by glycogenesis or β-glycogen formation during early differentiation (stage II), i.e. before the secretory activity, which is predominantly protein synthesis for shell-globule formation (stage III); (ii) the very heavy accumulation of large lipid droplets during the final stage of cytodifferentiation (stage IV); and (iii) the small number of β-glycogen particles present in mature vitellocytes. Ultracytochemical staining with PA-TCH-SP for glycogen proved positive for a small number of β-glycogen particles in differentiating and mature vitellocytes. Hypotheses, concerning the interrelationships of patterns of vitellogenesis, possible modes of egg formation, embryonic development and life-cycles, are commented upon.     

Differentiation of cytoplasmic organelles and storage of yolk during vitellogenesis in Hemidiaptomus ingens and Mixodiaptomus kupelwieseri (Copepoda,Calanoida)

These investigations concern two freshwater calanoid copepods Hemidiaptomus ingens and Mixodiaptomus kupelwieseri. The first aspect of the research relates to the processes involved in the formation and the differentiation of the ooplasmic organelles at the time of primary vitellogenesis. During this phase, a number of complex associations develop in the ooplasm. They consist chiefly of nuage-like structures, corresponding to extruded nuclear material, and vesicular formations, some arising from the nuclear envelope and the others neoformed in the ooplasm. These associations represent centers of maturation for ribosomes and synthesis for reticulum membranes. Annulate lamellae may be observed near these associations. Biogenesis of the reticulum always precedes the differentiation of the Golgi apparatus. Indeed, the dictyo-somes develop in characteristic complexes including endoplasmic reticulum cisternae and numerous vesicles resulting from intensive blebbing from cisternae. The second aspect of this research concerns yolk synthesis and accumulation of hyaloplasmic inclusions. A preliminary synthesis of yolk occurs early in these complexes and becomes more important after achievement of Golgi apparatus biogenesis. However, the most important yolk storage results from exogenous molecules and consists of complex globules, which develop into the ooplasm during secondary vitellogenesis. Formation of these globules is associated with the accumulation of two categories of inclusions in the hyaloplasm, i.e., lipid droplets and clusters of glycogen particles. At the end of vitellogenesis, a new type of endogenous material develops into small cisternae localized in the cortical ooplasm. © 1993 Wiley-Liss, Inc.     

Ultrastructural study of vitellogenesis and oogenesis of Metadena depressa (Stossich, 1883) Linton, 1910 (Digenea,Cryptogonimidae), intestinal parasite of Dentex dentex (Pisces,Teleostei)

The ultrastructural organization of the female reproductive system of Metadena depressa, digenean intestinal parasite of Sparidae (Dentex dentex), was investigated by electron microscopy. The vitellogenesis is divided into four stages: stage I, vitellocytes have a cytoplasm mainly filled with ribosomes and few mitochondria; stage II, beginning of the synthetic activity; stage III, active shell globule clusters synthesis; stage IV, mature vitellocytes are filled with shell globule clusters and generally contain several large lipid droplets. Glycogen granules are grouped at the periphery of the cell. The three stages of the oogenesis process take place in the ovary: stage I, oogonia are undifferentiated small cells located at the periphery of the organ; stage II, primary oocytes possess a higher nucleo-cytoplasmic ratio and a nucleus with a nucleolus and synaptonemal complexes indicating the zygotene-pachytene stage of the first meiotic division; stage III, mature oocytes are located in the proximal region of the organ and possess a cytoplasmic chromatoid body and cortical granules in a monolayer close to the periphery of the cell.     

Ultrastructure of the vitelline follicles of Gorgoderina vitelliloba (Trematoda: Gorgoderidae)

An electron microscope study of the vitelline follicles of Gorgoderina vitelliloba indicates that they contain vitelline cells in various stages of development. Juvenile cells are small and characterised by a little cytoplasm. During differentiation a large amount of granular endoplasmic reticulum develops. In more mature cells, indistinct Golgi complexes give rise to globules of shell protein which migrate to form clusters at the periphery of the cell. Further maturation results in the appearance of large lipid bodies in the vitelline cell cytoplasm.Developing vitelline cells are ensheathed by nurse cell cytoplasm containing numerous small vacuoles which appear to be derived from smooth endoplasmic reticulum. It is suggested that nurse cells may have a role in selection and transport of nutrient material for vitelline cells and that they manufacture precursors of lipid which is subsequently stored as a food reserve in mature vitelline cells. Possible transport sites between parenchymal cells and nurse cells were identified.     

Fine structure of the fat body in the female tickOrnithodoros (Pavlovskyella) erraticus (Ixodoidea: Argasidae)

The fat body in femalOrnithodoros (Pavlovskyella) erraticus Lucas consists of cell strands slung in the haemolymph but abundant around the tracheal trunks, midgut and reproductive system. The narrow cisternae of rough endoplasmic reticulum (RER) occupying most of the cell cytoplasm develop rapidly after feeding and become responsible for the formation of lipid droplets associated with glycogen particles. During oogenesis, RER and Golgi bodies are involved in protein synthesis which could be related to vitellogenesis. No ultrastructural investigations appear to have been made on the fat body in argasid ticks.     

An ultrastructural study of oogenesis in a marine triclad

The ultrastructural features of oocyte differentiation were studied in the marine triclad Cercyra hastata. Oocytes at several stages of maturation, each surrounded by follicle cell projections, are present within each of the two ovaries. A pre-vitellogenic and a vitellogenic stage have been detected in the oogenesis of C. hastata. The pre-vitellogenic stage is mainly characterized by an increase in the nuclear and nucleolar volume and activity, and the appearance and development of cortical granule precursors which are elaborated by the Golgi complex. In early phases of the vitellogenic stage, intense delamination and blebbing of the nuclear envelope occurs which probably contributes to an increase in number of cytoplasmic membranes and to transfer of nuclear material to the cytoplasm. The rough endoplasmic reticulum is extensively developed and often assumes a ‘whorl’ array. Several areas of yolk precursor formation appear in the whorls. Numerous 2–5 μm protein yolk globules are subsequently formed which appear surrounded by a double membrane (cisternae of the smooth endoplasmic reticulum) and become randomly distributed throughout the cytoplasm of mature oocytes. The peripheral ooplasm is occupied by a monolayer of electron-dense cortical granules. Finally, the evolutionary significance of the autosynthetic mechanism of yolk production is discussed.     

Morphology and ultrastructure of the adult ovarian cycle in Mithracidae (Crustacea,Decapoda, Brachyura,Majoidea)

The ultrastructure of the ovary during development and yolk production is poorly known in Brachyura and Majoidea in particular. Here, we describe the histology, histochemistry and ultrastructure of the adult ovarian cycle in four Mithracidae species from three different genera: Mithrax hispidus, Mithrax tortugae, Mithraculus forceps and Omalacantha bicornuta. All species showed a similar pattern of ovarian development and vitellogenesis. Macroscopically, we detected three stages of ovarian development: rudimentary (RUD), developing (DE) and mature (MAT); however, in histological and ultrastructural analyses, we identified four stages of development. The oocytes of the RUD stage, during endogenous vitellogenesis, have basophilic cytoplasm filled with dilated rough endoplasmic reticulum. The reticulum lumen showed many granular to electron-dense materials among the different stages of development. The Golgi complexes were only observed in the RUD stage and are responsible for releasing vesicles that merge to the endogenous or immature yolk vesicles. At the early DE stage, the oolemma showed many coated and endocytic vesicles at the cortex. The endocytic vesicles merge with the endogenous yolk to form the exogenous or mature yolk vesicles, always surrounded by a membrane, characterizing exogenous vitellogenesis. The exogenous yolk vesicles comprise glycoproteins, showing only neutral polysaccharides. At the late DE stage, endocytosis still occurs, but the amount of endogenous yolk decreases while the exogenous yolk increases. The late DE stage is characterized by the beginning of chorion production among the microvilli. The MAT stage is similar to the late DE, but the endogenous yolk is restricted to a few cytoplasmic areas, the ooplasma is filled with exogenous yolk, and the oolemma has very few coated vesicles. In the MAT stage, the chorion is fully formed and shows two electron-dense layers. The ovarian development of the species studied has many similarities with the very little known Majoidea in terms of the composition, arrangement and increment of the yolk vesicles during oocyte maturation. The main differences are in the vitellogenesis process, where immature yolk formation occurs without the direct participation of the mitochondria but with the participation of the rough endoplasmic reticulum in the endogenous phase.     

Fine Structure of the Ovarian Development in the Orb-web Spider, Nephila clavata

ABSTRACT Fine structural changes of the ovary and cellular composition of oocyte with respect to ovarian development in the orb-web spider, Nephila clavata were examined by scanning and transmission electron microscopy. Unlike the other arthropods, the ovary of this spider has only two kinds of cells-follicle cells and oocytes. During the ovarian maturation, each oocyte bulges into the body cavity and attaches to surface of the elongated ovarian epithelium through its peculiar short stalk attachments. In the cytoplasm of the developing oocyte two main types of yolk granules, electron-dense proteid yolk and electron-lucent lipid yolk granules, are compactly aggregated with numerous glycogen particles. The cytoplasm of the developing oocyte contains a lot of ribosomes, poorly developed rough endoplasmic reticulum, mitochondria and lipid droplets. These cell organelles, however, gradually degenerate by the later stage of vitellogenesis. During the active vitellogenesis stage, the proteid yolk is very rapidly formed and the oocyte increases in size. However, the micropinocytosis invagination or pinocytotic vesicles can scarcely be recognized, although the microvilli can be found in some space between the oocyte and ovarian epithelium. During the vitellogenesis, the lipid droplets in the cytoplasm of oocytes increase in number, and become abundant in the peripheral cytoplasm close to the stalks. On completion of the yolk formation the vitelline membrane, which is composed of an inner homogeneous electron-lucent component and an outer layer of electron-dense component is formed around the oocyte.     

PRODUCTION OF MEMBRANE WHORLS IN RAT LIVER BY SOME INHIBITORS OF PROTEIN SYNTHESIS

Inhibitors of protein synthesis capable of differential effects on nascent peptide synthesis on membrane-bound and free polyribosomes were employed to investigate the structure and function of cellular membranes of liver. The formation of membranous whorls in the cytoplasm and distension of nuclear membranes were induced by inhibitors of protein synthesis (i.e., cycloheximide and emetine) which predominantly interfere with nascent peptide synthesis on membrane-bound polyribosomes in situ. Other inhibitors of protein synthesis such as puromycin and fusidic acid, which inhibit nascent peptide synthesis on both free and membrane-bound polyribosomes, and chloramphenicol, which inhibits mitochondrial protein synthesis, did not induce these alterations. Cycloheximide, puromycin, and chloramphenicol produce some common cellular lesions as reflected by similar alterations in morphology, such as swelling of mitochondria, degranulation of rough endoplasmic reticulum, and aggregation of free ribosomes. The process of whorl formation in the cytoplasm, the incorporation of [³H]leucine and of [³H]choline into endoplasmic reticulum and the total NADPH-cytochrome c reductase activity of the endoplasmic reticulum were determined. During maximum formation of membranous whorls, [³H]leucine incorporation into cytoplasmic membranes was inhibited, while [³H]choline incorporation into these structures was increased; maximum inhibition of protein synthesis and stimulation of choline incorporation into endoplasmic reticulum, however, preceded whorl formation. Cycloheximide decreased the activity of NADPH-cytochrome c reductase of rough endoplasmic reticulum, but increased NADPH-cytochrome c reductase activity of smooth endoplasmic reticulum. In addition, cycloheximide decreased the content of hemoprotein in both the microsomal and mitochondrial fractions of rat liver, and the activities of mixed function oxidase and of oxidative phosphorylation were impaired to different degrees. Succinate-stimulated microsomal oxidation was also inhibited. The possible mechanisms involved in the formation of membranous whorls, as well as their functions, are discussed.     

Ultrastructural features of the nucleus and cytoplasm of rat trophoblast cells of the connective zone of the placenta and labyrinth

Ultrastructural organization of the rat trophoblast cells in the connective zone of placenta and labyrinth was investigated on the 12-14th days of gestation. A clear distinction was revealed in the cytoplasm ultrastructure of two cell subpopulations within the connective zone of placenta, i.e. glycogen and trophospongium cells. The former display a well defined network of long thin channels of granular endoplasmic reticulum situated mainly around the glycogen clusters. On the contrary, the latter are rich in the smooth endoplasmic reticulum but lacking glycogen accumulation. Differences in the nucleolar ultrastructure in these two cell subpopulations are not very considerable. A characteristic feature of glycogen cells is the presence of numerous round or oval small-fibrillar nucleolus-like bodies with the diameter of granules 20 nm. The trophoblast cells of the labyrinth are heavily laden with polysomes, which sometimes attach to short channels of the granular endoplasmic reticulum. Not often there occur short profiles of the agranular endoplasmic reticulum. Nucleolus-like bodies are found in all the cell types examined. This means that the nucleolus-like bodies may arise not only on the lampbrush chromosomes in the oocytes or polytene chromosomes, but also in the somatic cells which are capable of dividing mitotically.     

A fine structural study of cytodifferentiation during cleavage, blastula, and gastrula stages of Fundulus heteroclitus

The fine structure of cleavage, blastula, and gastrula stages of Fundulus heteroclitus was investigated. Cleavage blastomeres are relatively unspecialized, containing few or poorly developed organelles. Beginning in blastula stages, signs of differentiation were noted, including development of the endoplasmic reticulum and Golgi apparatus and appearance of a primary nucleolus and polyribosomes. More extensive structural specializations occur in gastrula stages, including further development of the endoplasmic reticulum and appearance of a granular component in the nucleolus. These changes are associated with cell differentiation and an increased capacity for protein synthesis, and may be preparatory to subsequent histogenesis. The periblast is a continuous syncytial cytoplasmic layer located between the blastodisc and yolk and is formed during late cleavage by incomplete division of the cytoplasm of the blastodisc. Cytoplasmic projections extend from the periblast (and from the basal region of cleavage blastomeres prior to formation of the periblast) into the yolk and function in uptake of yolk material in the absence of pinocytosis. Yolk material appears to be digested by the periblast and transferred into the segmentation cavity where it is available to the blastomeres. Protein granules, lipid droplets, glycogen, crystalline arrays, and multivesicular bodies are related to food storage and utilization by blastomeres. The yolk gel layer enclosing the yolk sphere was found to be a thin layer of cytoplasm continuous with the margin of the periblast and is renamed the yolk cytoplasmic layer.     

Observations on the role of smooth endoplasmic reticulum in glucocorticoid-induced hepatic glycogen deposition

We have studied by quantitative electron microscopy the relationship of specific hepatic cellular organelles to glycogen synthesis using dexamethasone, a potent synthetic glucocorticoid, to induce glycogen deposition in livers of adrenalectomized rats. Chemical and ultrastructural glycogen determinations revealed that the livers of fasted adrenalectomized rats had very low glycogen levels. Dexamethasone caused a time-related increase in hepatic glycogen which was the result of increases in the number of hepatocytes depositing glycogen and the amount of glycogen in each cell. The surface density of smooth endoplasmic reticulum (SER) in centrilobular and periportal hepatocytes also increased after treatment with dexamethasone; this increase preceded glycogen deposition. The newly deposited glycogen was spatially associated with membranes of SER, and a continued increase in SER surface density was correlated temporally with the increasing glycogen volume density. In both centrilobular and periportal hepatocytes, the suface density of rough endoplasmic reticulum (RER) initially decreased after dexamethasone administration but later increased. These data support the hypothesis that dexamethasone-induced enhancement of SER is functionally associated with the increase in glycogen, and that although the initial increase in SER may occur through transformation of RER to SER, later increases in SER require synthesis of new membranes.     

东方杯叶吸虫卵黄腺和卵巢的超微结构研究

本文应用透射电镜观察了东方杯叶吸虫卵黄腺和卵巢的超微结构,并与体外培养成虫进行比较。根据形态特征和内含物的存在情况,将卵黄细胞和卵母细胞的发育均分为不同时期,详细描述了各期的形态特征,探讨了卵黄球和皮质颗粒等内含物的生理功能。体外培养成虫成熟卵黄细胞中有散在的卵黄物质,成熟卵母细胞中线粒体囊泡化,这些可作为体外培养的评价指标。     

Morphometric analysis of hepatocytes from rats subjected to compound 48/80-induced anaphylactic shock

Morphometric and biochemical techniques were used to analyze hepatic glycogen, endoplasmic reticulum, and mitochondrial matrix granules in rats treated with compound 48/80 to induce an anaphylactic-like state of shock. Thirty minutes after insult there was a significant decrease in glycogen and mitochondrial matrix granules, an increase in rough endoplasmic reticulum (RER), and no change in smooth endoplasmic reticulum (SER). Less glycogen in experimental rats substantiated a previously described glycogenolytic response to compound 48/80. The decrease in matrix granules implies a loss and/or shift in intramitochondrial calcium as occurs in epinephrine-induced glycogenolysis in the rat. Since other glycogenolytic agents, e.g. glucagon, and starvation stimulate an increase in SER presumably from RER, the present morphological data suggest the increase in RER may precede proliferation of SER from RER.     

Viral Glycoproteins Accumulate in Newly Formed Annulate Lamellae following Infection of Lymphoid Cells by Human Herpesvirus 6

Ultrastructural analysis of HSB-2 T-lymphoid cells and human cord blood mononuclear cells infected with human herpesvirus 6 revealed the presence, in the cell cytoplasm, of annulate lamellae (AL), which were absent in uninfected cells. Time course analysis of the appearance of AL following viral infection showed that no AL were visible within the first 72 h postinfection and that their formation correlated with the expression of the late viral glycoprotein gp116. The requirement of active viral replication for AL neoformation was further confirmed by experiments using inactivated virus or performed in presence of the viral DNA polymerase inhibitor phosphonoacetic acid. Both conventional electron microscopic examination and immunogold fracture labeling with anti-endoplasmic reticulum antibodies indicated a close relationship of AL with the endoplasmic reticulum and nuclear membranes. However, when the freeze-fractured cells were immunogold labeled with an anti-gp116 monoclonal antibody, AL membranes were densely labeled, whereas nuclear membranes and endoplasmic reticulum cisternae appeared virtually unlabeled, showing that viral envelope glycoproteins selectively accumulate in AL. In addition, gold labeling with Helix pomatia lectin and wheat germ agglutinin indicated that AL cisternae, similar to cis-Golgi membranes, contain intermediate, but not terminal, forms of glycoconjugates. Taken together, these results suggest that in this cell-virus system, AL function as a viral glycoprotein storage compartment and as a putative site of O-glycosylation.     

Development of secretory activity in the seminal vesicle of the male migratory grasshopper,Melanoplus sanguinipes (fabr.) (Orthoptera : Acrididae)

This paper describes the ultrastructure of the seminal vesicle and the isoelectric focusing patterns of its secretion during sexual maturation and after allatectomy in Melanoplus sanguinipes (Fabr.) (Orthoptera : Acrididae). In epithelia from seminal vesicles of newly fledged males, the rough endoplasmic reticulum is well developed, and Golgi complexes are elaborate, which indicates the gland is metabolically active. The cells also contain large glycogen deposits and the lumen microvilli are well differentiated. These ultrastructural features are more dominant in 24-hr-old adults where the cytoplasm is clearly differentiated into basal and apical regions. Basally, the cytoplasm is dominated by rough endoplasmic reticulum, large Golgi complexes, glycogen deposits and numerous mitochondria, while the apical cytoplasm is filled with large secretory and/or lysosomal vesicles. Between days 3 and 7, the ultrastructural features change little other than the rough endoplasmic reticulum cisternae, which become vesicular. Analysis by isoelectric focusing shows that the amount of secretory protein increases with age until day 3, at which time the gland contains its full complement of secretion. In seminal vesicles from allatectomized insects, ultrastructural features of cells and isoelectric focusing patterns of the secretion arc identical to those from normal males.     

Development of a nuclear polyhedrosis virus in midgut cells and penetration of the virus into the hemocoel of the armyworm,Pseudaletia unipuncta

The development of a nuclear polyhedrosis virus (NPV) in larval midgut cells of the armyworm, Pseudaletia unipuncta, is similar to that of other NPV. In the nucleus, the envelopes around the nucleocapsids seem to be derived de novo or from the inner layer of the nuclear envelope wich forms cisternae, blebs, or infoldings. The nucleocapsids are also enveloped by synhymenosis during passage through the nuclear membrane, the cell membrane, or the endoplasmic reticulum membrane. Both enveloped and unenveloped nucleocapsids may enter the cytoplasm through the nuclear pore or budding through the nuclear membrane. From the cytoplasm the virions may enter the hemocoel through the basal cell and basement membranes or through the endoplasmic reticulum, intercellular space, and the basement membrane.