Ultrastructure of the tegument of the cestode Paraechinophallus japonicus (Bothriocephalidea: Echinophallidae), a parasite of the bathypelagic fish Psenopsis anomala

Abstract. The ultrastructure of the tegument in Paraechinophallus japonicus (Bothriocephalidea: Echinophallidae), a cestode parasite of the bathypelagic fish Psenopsis anomala , was studied using scanning and transmission electron microscopy. Paraechinophallus japonicus lacks a true scolex. Four different types of microtriches have been observed on the tegumental surface of P. japonicus. Capilliform (∼2.3-μm long) and blade-like spiniform (∼1.4-μm long) microtriches are intermingled on the surface of the pseudoscolex. Capilliform microtriches are distinct in possessing a short base and a long electron-lucent cap. The strobila is covered with two types of microtriches, namely filiform (∼2.1-μm long) and tusk-shaped microtriches (≤4.5-μm long). Tusk-shaped microtriches are limited to the posterior border of each proglottid and are characterized by a short and narrow base, and a large and wide, sharply pointed, electron-dense cap. Similar tusk-shaped microtriches were previously found in members of the family Echinophallidae and may represent an autapomorphy of echinophallid cestodes, all of them being parasitic in centrolophid fish. A unified terminology of microthrix parts is proposed.     

A thin-section and freeze-fracture study of microfilament-membrane attachments in choroid plexus and intestinal microvilli

Choroid plexus and intestinal microvilli in thin sections have microfilaments in the cytoplasm adjacent to the membranes, and in replicas have broken strands of filaments in both cytoplasm and on E faces of plasm membranes. The microfilaments contain actin as indicated by their binding of heavy meromyosin (HMM). In sections of choroid plexus, the microfilaments are 7-8 nm in diameter and form a loose meshwork which lies parallel to the membrane and which is connected to the membranes both by short, connecting filaments (8 times 30 nm) and dense globules (approximately 15-20 nm). The filamentous strands seen in replicas are approximately 8 nm in diameter. Because they are similar in diameter and are connected to the membrane, these filamentous strands seen in replicas apparently represent the connecting structures, portions of the microfilaments, or both. The filamentous strands attached to the membrane are usually associated with the E face and appear to be pulled through the P half-membrane. In replicas of intestinal brush border microvilli, the connecting strands attaching core microfilaments to the membrane are readily visualized. In contrast, regions of attachment of core microfilaments to dense material at the tips of microvilli are associated with few particles on P faces and with few filamentous strands on the E faces of the membranes. Freeze-fracture replicas suggest a morphologically similar type of connecting strand attachment for microfilament-membrane binding in both choroid plexus and intestinal microvilli, despite the lack of a prominent core bundle of microfilaments in choroid plexus microvilli.     

Surface ultrastructure of the elasmobranchia parasitizing <Emphasis Type="Italic">Grillotiella exilis</Emphasis> and <Emphasis Type="Italic">Pseudonybelinia odontacantha</Emphasis> (Trypanorhyncha,Cestoda)

The surface ultrastructure of two monotypic trypanorhynch genera is described based on new material of Grillotiella exilis (Linton, 1909) and type material of Pseudonybelinia odontacantha Dollfus 1966. In G. exilis, spiniform microtriches cover the bothrial surfaces and the anterior part of the pars vaginalis posterior to the bothria. Bifurcate microtriches adorn the bothrial margins, filiform microtriches the scolex peduncle, and capilliform microtriches the posterior scolex end. This microthrix pattern resembles that found in, e.g., Grillotia erinaceus (van Beneden, 1858), with the difference that the anterior part of the pars vaginalis is covered with a collar of multidigitate palmate microtriches. The position of Grillotiella within the Grillotiinae, Lacistorhynchidae is supported based on these data. The bothria and scolex peduncle of P. odontacantha are covered with acerosate and unciniform microtriches on the distal bothrial surface and capilliform microtriches on the scolex peduncle. Short filiform microtriches cover the appendix. The microthrix pattern resembles that of the Tentaculariidae but with unciniform and acerosate microtriches densely covering the entire distal bothrial surface. Tegumental grooves are present on the posterior bothrial margin. They can be distinguished from bothrial pits in otobothrioid trypanorhynchs in having similar unciniform microtriches compared to the other parts of the bothrial surface and in lacking any spiniform microtriches. With the absence of bothrial pits as characteristic for the otobothrioids and its characteristic microthrix pattern, P. odontacantha together with Paranybelinia otobothrioides Dollfus 1966, both belonging to the Paranybeliniidae change their position in the most recent system from the Otobothrioidea into the Tentacularioidea.     

Ultrastructure of the midgut and the adhering tubular salivary glands of Frankliniella occidentalis (Pergande) (Thysanoptera : Thripidae)

The ultrastructure of the midgut and the tubular salivary glands of Frankliniella occidentalis (Thysanoptera : Thripidae) is described. The microvilli have 2 different types of glycocalyx: in the anterior part of the midgut they are surrounded by a myelin-like membrane; in the posterior region, the microvilli have numerous rod-like projections arranged to form a continuous layer. Microfilaments longitudinally cross each microvillus; the microfilaments contain F-actin. Tubular salivary glands flank the midgut but do not fuse with it. The distal part of these glands have microvillated cells containing large amounts of electron-transparent material. The cells of the proximal part are lined with a thin cuticle.     

Brush-border formation in the midgut of an insect,Calliphora erythrocephala meigen

Summary The embryonic development of the brush-border of anterior midgut cells of Calliphora was studied by electron microscopy. Dense surface-forming vesicles, as described by Bonneville (1970), are found prior to microvillus formation. These dense vesicles provide membranous and coating material for the moulding of the microvilli. The number of dense vesicles increases rapidly to a maximum just before brush-border formation, after which it decreases very rapidly, accompanied by an increase in the number of microvilli. Formation of microvilli proceeds in essentially the same way as in Xenopus. First, some of the vesicles fuse with the apical cell membrane, resulting in an increase of the cell surface, part of which is coated with filamentous material deriving from the dense vesicles. This in turn leads to bulging, and short irregular microvilli appear. These are erected and elongated.Prefabricated tubular elements are believed to play a part in this erection and elongation, probably due to the unwinding of spirally coiled strands.Microvillus formation proper lasts 2 to 3 hours in Calliphora. Almost the entire amount of membranous and coating material is prefabricated prior to the formation of microvilli.     

The actin microfilament network within elongating pollen tubes of the gymnospermPicea abies (Norway spruce)

Summary Actin microfilaments form a dense network within pollen tubes of the gymnosperm Norway spruce (Picea abies). Microfilaments emanate from within the pollen grain and form long, branching arrays passing through the aperture and down the length of the pollen tube to the tip. Pollen tubes are densely packed with large amyloplasts, which are surrounded by branching microfilament bundles. The vegetative nucleus is suspended within the elongating pollen tube within a complex array of microfilaments oriented both parallel to and perpendicular with the growing axis. Microfilament bundles branch out along the nuclear surface, and some filaments terminate on or emanate from the surface. Microfilaments in the pollen tube tip form a 6 m thick, dense, uniform layer beneath the plasma membrane. This layer ensheathes an actin depleted core which contains cytoplasm and organelles, including small amyloplasts, and extends back 36 m from the tip. Behind the core region, the distinct actin layer is absent as microfilaments are present throughout the pollen tube. Organelle zonation is not always maintained in these conifer pollen tubes. Large amyloplasts will fill the pollen tube up to the growing tip, while the distinct layer of microfilaments and cytoplasm beneath the plasma membrane is maintained. The distinctive microfilament arrangement in the pollen tube tips of this conifer is similar to that seen in tip growth in fungi, ferns and mosses, but has not been reported previously in seed plants.     

An ultrastructural analysis of mature sperm from the crayfish Pacifastacus leniusculus,Dana

Sperm from the crayfish, Pacifastacus leniusculus, resemble other reptantian sperm in that they are composed of an acrosome, subacrosomal region, nucleus, membrane lamellar complex, and spikes which radiate from the nuclear compartment. The acrosome (PAS positive vesicle) can be subdivided into three regions: the apical cap, crystalline inner acrosomal material, and outer acrosomal material which is homogeneous except for a peripheral electron dense band. The nucleus contains uncondensed chromatin and bundles of microtubules which project into the spikes. The orientation of the microtubule bundles relative to the nuclear envelope near the base of the subacrosomal region suggests that the nuclear envelope may function in the organization of the spike microtubules.     

Smooth muscle and myofibroblast-like cells in the perimedullary stromal basket of kidneys from ringed seals (Phoca hispida)

Summary The medullary pyramid of renculi in kidneys of ringed seals (Phoca hispida) is enclosed by a basket composed of ribbons of stromal tissue continuous with the wall of the calyx. Branched smooth muscle cells with well-developed Golgi complexes and rough endoplasmic reticulum and only an incomplete external lamina are the principal cells in sites near the origin of the ribbons from the calycal wall. Deeper in the corticomedullary junctional region, smooth muscle is progressively replaced with stellate or spindle-shaped cells exhibiting structural characteristics intermediate between those of fibroblasts and smooth muscle fibers. These myofibroblast-like cells contain arrays of parallel microfilaments 6–8 nm thick with associated focal densities and subplasmalemmal dense plaques, caveolae, elongate, often deeply wrinkled nuclei, and well-developed Golgi complexes and rough endoplasmic reticulum. Material resembling external lamina is associated with parts of the surfaces of most myofibroblast-like cells and intermediate junctions are present. Fibroblasts lacking arrays of parallel microfilaments are a minority at any level in the stromal ribbons. Interstitial cells in the vicinity of the corticomedullary junction show similar myofibroblast-like characteristics. The smooth muscle and myofibroblast-like cells presumably assist expression of urine from the papilla and calyx, and possibly participate as pacemakers for the urinary tract.     

Follicular placenta of the viviparous fish,Heterandria formosa: II. Ultrastructure and development of the follicular epithelium

Embryos of the viviparous poeciliid fish, Heterandria formosa, develop to term in the ovarian follicle where they undergo a 3,900% increase in embryonic dry weight. Maternal-embryonic nutrient transfer occurs across a follicular placenta that is formed by close apposition of the embryonic surface (i.e., the entire body surface during early gestation and the pericardial amnionserosa during mid-late gestation) to the follicular epithelium. To complement our recent study of the embryonic component of the follicular placenta, we now describe the development and fine structure of the maternal component of the follicular placenta. Transmission electron microscopy reveals that the ultrastructure of the egg envelope and the follicular epithelium that invests vitellogenic oocytes is typical of that described for teleosts. The egg envelope is a dense matrix, penetrated by microvilli of the oocyte. The follicular epithelium consists of a single layer of cuboidal cells that lack apical microvilli, basal surface specializations, and junctional complexes. Follicle cells investing the youngest embryonic stage examined (Tavolga's and Rugh's stage 5–7 for Xiphophorus maculatus) also lack apical microvilli and basal specializations, but possess junctional complexes. In contrast, follicle cells that invest embryos at stage 10 and later display ultrastructural features characteristic of transporting epithelial cells. Apical microvilli and surface invaginations are present. The basal surface is extensively folded. Apical and basal coated pits are present. The cytoplasm contains a rough endoplasmic reticulum, Golgi complexes, and dense staining vesicles that appear to be lysosomes. The presence of numerous apically located electron-lucent vesicles that appear to be derived from the apical surface further suggests that these follicle cells may absorb and process follicular fluid. The egg envelope, which remains intact throughout gestation and lacks perforations, becomes progressively thinner and less dense as gestation proceeds. We postulate that these ultrastructural features, which are not present in the follicles of the lecithotrophic poeciliid, Poecilia reticulata, are specializations for maternal-embryonic nutrient transfer and that the egg envelope, follicular epithelium, and underlying capillary network form the maternal component of the follicular placenta. © 1994 Wiley-Liss, Inc.     

Comparison of the localization of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Cry1A δ-endotoxins and their binding proteins in larval midgut of tobacco hornworm, <Emphasis Type="Italic">Manduca sexta</Emphasis>

Tobacco hornworm, Manduca sexta, is a model insect for studying the action of Bacillus thuringiensis (Bt) Cry toxins on lepidopterans. The proteins, which bind Bt toxins to midgut epithelial cells, are key factors involved in the insecticidal functions of the toxins. Three Cry1A-binding proteins, viz., aminopeptidase N (APN), the cadherin-like Bt-R₁, and membrane-type alkaline phosphatase (m-ALP), were localized, by immunohistochemistry, in sections from the anterior, middle, and posterior regions of the midgut from second instar M. sexta larvae. Both APN and m-ALP were distributed predominantly along microvilli in the posterior region and to a lesser extent on the apical tip of microvilli in the anterior and middle regions. Bt-R₁ was localized at the base of microvilli in the anterior region, over the entire microvilli in the middle region, and at both the apex and base of microvilli in the posterior region. The localization of rhodamine-labeled Cry1Aa, Cry1Ab, and Cry1Ac binding was determined on sections from the same midgut regions. Cry1Aa and Cry1Ab bound to the apical tip of microvilli almost equally in all midgut regions. Binding of Cry1Ac was much stronger in the posterior region than in the anterior and middle regions. Thus, binding sites for Bt proteins and Cry1A toxins are co-localized on the microvilli of M. sexta midgut epithelial cells.     

Novel surface specialization on a sea anemone egg: “Spires” of actin-filled microvilli

The ultrastructure of the “spiny” surface of Tealia crassicornis eggs is examined in detail by scanning and transmission electron microscopy in order to understand its function. Long microvilli are clustered together in spiral aggregates of 50–75 microvilli called “spires.” There are about 15,000 spires per egg. Dense bundles of microfilaments making up the cores of these microvilli are shown to be composed of actin by staining with the fluorescent dye nitrobenzoxadiazole (NBD)-phallacidin. It is postulated that the bundles of actin and the spires of microvilli are stiff and provide reinforcement to the egg surface. Such postulated properties would provide physical protection for these large eggs which, unlike the eggs of most invertebrates, appear to lack all extracellular investing coats.     

A new morphological aspect of the brush cells of the mouse gallbladder epithelium

Summary The brush cells of the gallbladder epithelium of the mouse have microvilli not only at their luminal border but also on their lateral surface, from the level of the nucleus to the junctional complex. The lateral microvilli radiate from the brush cell in all directions, contain a core of filaments, and penetrate up to 3 m into the adjacent cells. The microvilli in these locations display small desmosomes at their base.     

Isolation of microvillar microfilaments and associated transmembrane complex from ascites tumor cell microvilli

The association of microvillar microfilaments with the microvillar membrane actin-containing transmembrane complex of MAT-C1 13762 ascites tumor cell microvilli has been investigated by differential centrifugation, gel electrophoresis and electron microscopy of detergent extracts of the isolated microvilli. Several methods have been used to reduce breakdown and solubilization of the microfilament core actin during the detergent extractions for preparation of microvillar core microfilaments. Gel electrophoresis of differential centrifugation fractions demonstrated that over 70% of the total microvillus actin could be pelleted with microfilament cores at 10 000 g under extraction conditions which reduce filament breakdown. Transmission electron microscopy (TEM) of all of the core preparations showed arrays of microfilaments and small microfilament bundles. The major protein components of the microfilament cores, observed by sodium dodecyl sulfate (SDS) electrophoresis, were actin and alpha-actinin. Among the less prominent polypeptide components was a 58 000 Dalton polypeptide (58 K), previously identified as a member of the MAT-Cl transmembrane complex. This three-component complex contains, in addition to 58 K, actin associated directly and stably with a cell surface glycoprotein (Carraway, CAC, Jung, G & Carraway, K L, Proc. natl acad. sci. US 80 (1983) 430). Evidence that the apparent association of complex with the microfilament core was not due simply to co-sedimentation was provided by myosin affinity precipitation. These results provide further evidence that the transmembrane complex is a site for the interaction of microfilaments with the microvillar plasma membrane.     

Ultrastructure of the epidermis in the ice worm, Mesenchytraeus solifugus

The ice worm is adapted for life at O°C. A survey of the ultrastructure of the cuticle, epidermal epithelium and basement membrane does not reveal any features which self-evidently correlate with such metabolic specialization; instead, these tissues are much like those of the earthworm and some freshwater oligochaetes. The cuticular fibers are unstriated. Epithelial cells aresuggested as the source of cuticular material. Epithelial microvilli penetrate the cuticle. There is an array of membrane bound bodies on the cuticle surface. The basement membrane fibers are transversely striated and are oriented in crossed lamellae. The junctional complex is represented by azonula adhaerens and septate desmosome.     

The ultrastructure of the cuticle of some British lumbricids (Annelida)

The ultrastructure of the cuticle of 11 species of British lumbricids is described. The number of unbanded collagenous fibre layers varies with the species and is roughly proportional to the size of the adult worm. Four zones are discernible in the cuticle matrix in all species except E. foetida where the outermost zone shows subdivision.
Microvilli, cytoplasmic extensions from the epithelial surface, occur. The "long" and "short" microvilli of other workers are shown to be different views of the same structure. The microvilli have an ovoid base with the two poles forming low shoulders on either side of the ascending microvillus. The bases are oriented at right-angles to the longitudinal axis of the worm and the microvilli are arranged in regular staggered rows along the same axis. Details of the spatial alignment of the microvilli are given and the possible role of these regularly arranged structures as factors in the orientation of the collagen fibre unit filaments is discussed, and it is speculated that they might also have some proprioceptive function.
Distally the microvilli terminate among the surface epicuticular projections and are shown to give rise to them. The epicuticular projections are peanut-shell shaped with a distinct substructure consisting of a 8 nm electron dense lining and two parallel dense discs, each 8 nm deep, above the "waist".
The mucous cell pores are lined with electron pale microfibrillar material and at the base of the pore a circlet of 13–15 short microvilli, with prominent tonofilaments, arises from the cytoplasm of the mucous cell. Surrounding the pore microvilli are numerous, small, membrane bound mucous pore particles.     

Fine structure and freeze-etch study of the protoscolex tegument of Echinococcus multilocularis (cestoda)

Freeze-etch replicas of the protoscolex tegument of Echinococcus multilocularis were examined and compared with conventional thin sections by TEM. The microtopography of the protoscolex tegument was also examined by SEM. The protoscolex consisted of morphologically-distinct, apical and basal tegumentary regions, the latter of which lacked microtriches. The hook area of the apical region contained long, slender, filamentous microtriches that obscured the hook arrangement. These microtriches were structurally different from those found on the suckers and rostellum of the protoscolex. Freeze-etch replicas of the tegumental membrane of the sucker and rostellar microtriches showed that the protoplasmic (P) and exoplasmic (E) faces of the microthrix base and tip contained numerous intramembranous particles (IMP). The densities of the IMP on both the P and E faces of the microthrix tip were approximately twice the number of the larger diameter IMP found on the P and E faces of the microthrix base. No freeze-etch replicas of the microtriches from the hook area were obtained. The basal tegumentary region of the protoscolex consisted of irregularly-distributed, knoblike processes that were variable in size and shape, and contained an electron-dense cap. The IMP on the P face of the knoblike processes measured approximately the same diameter as those on the P face of the microthrix base. However, their density was about half that of the latter. The density of IMP on the E face of the knoblike processes could not be determined from the freeze-etch replicas.     

The ultrastructure of the cornea-lens epidermis in the lateral eye of Limulus polyphemus

The structure of the “Corneagen,” i.e., the epidermis lying beneath the cornea-lens of the lateral eyes of the adult intermolt Limulus polyphemus was studied with light and electron microscopy. This layer is composed of heavily columnar cells containing a striking number of cytoplasmic microtubules. Many of the microtubules are grouped into compact bundles or fascicles, generally each cell having at least one microtubule bundle. The cornealens end of each cell has numerous microvilli, each with a core of delicate filaments. The crypts between microvilli end in extracellular expansions and plaques of electron dense amorphous material are associated with these terminal expansions. Cytoplasmic microtubules appear to insert into these dense areas. The basal ends of the cells are thrown into many pseudopodial processes which extend into the surrounding extracellular space. The cytoplasm of the pseudopodia is composed largely of microtubules and their associated low density halos. Junctional complexes consisting of zonulae adhaerens and septate desmosomes are present between adjacent cells. Mitochondria, ER, cytoplasmic vesicles, Golgi stacks and other ultrastructural details of the epidermal cells are described. The ultrastructure of a column of pigment free processes lying between the apex of the lens cone and the underlying photoreceptive portion of the ommatidium is also described. Ducts or vessels of uncertain origin are present in the inter-ommatidial spaces. Possible roles played by the microtubules, the significance of their disposition and of their association with the dense subsurface plaques are discussed in terms of intracellular support, epidermis-lens attachment and extracellular pattern determination. In addition, the likelihood of the dense plaques being the site of microtubule assembly is considered.     

Organization of the cross-filaments in intestinal microvilli

We studied the arrangement of the cross-filaments in intestinal microvilli to understand how microfilaments interact with the membrane. Observations on thin-sectioned or negatively stained microvilli with the electron microscope demonstrate that the cross-filaments on the core bundle lie opposite to one another and are spaced 32.5 nm apart. In sections grazing through the membranes, the cross-filaments appear as transverse stripes in a barber-polelike arrangement. The cross- filaments point away from the microvillus tip. This subfragments S1 or HMM. The cross filaments are associated not only with the microfilaments but also with electron-dense patches on the inside surface of the membrane. These results suggest the cross-filaments are arranged as a double helix around the core bundle. Furthermore, the cross-filaments can serve as in situ markers for microvillar polarity. Lastly, the cross-filaments interact not only with specific portions on the actin filaments but also with dense patches on the membrane. These observations are summarized in a model of the microvillus cytoskeleton.     

Ultrastructural investigation of the mechanism of muscle attachment to the gastropod shell

The ultrastructure of the muscle-shell attachment was investigated in the land pulmonate snails Helix aspersa, Anguispira altemata, in the freshwater pulmonate Laevipex sp., and in the freshwater prosobranch Pomacea paludosa. In all cases, a collagenous intercellular matrix and a specialized epithelium (tendon cells) intervene between the columellar muscle and the shell. These tendon cells are characterized by hemidesmosomes at both apical and basal ends, connected by thick bundles of microfilaments. The tendon cells do not insert into the shell directly by microvilli, as formerly thought, but by an extensive network of extracellular organic fibers.