Ultrastructure of the eyes of the larva of Neoheterocotyle rhinobatidis (Platyhelminthes, Monopisthocotylea), and phylogenetic implications.

The oncomiracidium of Neoheterocotyle rhinobatidis (Monogenea, Monopisthocotylea, Monocotylidae) has two pairs of eyes, each eye with a lens and pigment cup. The anterior eyes have a single rhadomere; the posterior ones, two rhadomeres. Lenses are part of the pigment cup cells, as indicated by cytoplasmic connections between them and the pigment cups, and they are of mitochondrial origin because mitochondrial cristae are present in the periphery of the lenses. This is the first time that mitochondrial lenses have been shown to exist in a neodermatan. Such lenses may be a synapomorphy of a large taxon comprising the Neodermata and its turbellarian sister groups, or they may have evolved convergently in several not closely related groups as a result of strong selection pressure to find a suitable habitat or host.     

Epidermis and protonephridia of a free-living platyhelminth, Mesocastrada führmanni Voltz, 1898 (Rhabdocoela, Typhloplanoida): An ultrastructural study

The ultrastructure of the epidermis and the protonephridia of the free-living rhabdocoel Mesoscastrada führmanni is described. The epidermis consists of polarized cells, the nucleus located in the basal part of the cell and the mitochondria in the apical part. The surface is entirely covered by cilia anchored in the cytoplasm by horizontal and vertical striated rootlets. Cilia of the flame bulbs also have horizontal and vertical striated rootlets. The weir apparatus of the cyrtocyte is composed of a single row of ribs connected by a thin “membrane” of extracellular material. Bundles of microtubules, located in the ribs originate in the centrioles. Epidermal cells and flame bulbs of M. führmanni closely resemble those of the other Typhloplanoida examined so far.     

The system of epidermal ciliary rootlets in Turbellaria

Summary The rootlets of the kinetic cilia form patterns of different types in the different turbellarian subgroups (cf. Rieger 1981). In the Acoela a rather complex system of ciliary rootlets is found in the epidermis (Dorey 1965; Hendelberg & Hedlund 1973; Bedini & Papi 1974). In the acoel Childia groenlandica (Levinsen) the four rootlets of each cilium make contact with those of adjacent cilia at two levels (Hendelberg & Hedlund 1974). Distinct granules are found in the interior of the main rootlets (Hendelberg & Hedlund 1974; Bedini & Papi 1974, Fig. 16) and basal bodies (Silveira 1972; Hendelberg & Hedlund 1974) of the epidermal kinetic cilia of acoels. Similar granules, probably of identical structure, can be seen in nemertodermatids, in the same positions (Tyler & Rieger 1977, Figs. 3 & 6). Such granules were studied in C. groenlandica with histochemical methods adapted for electron microscopy. Like Silveira (1972) I found the granules of the basal bodies to be Thiéry-positive, and thus evidently to be made up of or at least to contain polysaccharide material. The granules of the main rootlets were also found to be Thiéry-positive (Hendelberg 1976). Digestion experiments (Hendelberg & Hellmén 1978 and unpublished results) strongly support the concept that the granules are glycogen beta-particles.We know that cilia can function as kinetic organelles without any rootlets. But we are still uncertain about the function of the rootlets when occurring. Most probably they form an anchorage, a function which may be favoured by branching rootlets making contact with each other. Another function which has been discussed is the transmittance of impulses regulating the ciliary beat. Glycogen granules represent an energy deposit. The functional implication of these granules in the interior of the ciliary rootlets and basal bodies is not clear. However, the observations raise the question of how energy is transmitted to the cilia. Are the ciliary rootlets, when occurring, involved? This question will be further discussed, with references, in a future full report on the digestion experiments (to be published elsewhere).     

Ultrastructural study of the epidermis in two free-living Platyhelminthes, Mesostoma viaregginum and M. productum (Rhabdocoela, Typhloplanida)

Abstract. The epidermis of the free-living typhloplanids Mesostoma viaregginum and M. productum (Mesostominae) is described. In both species, the epidermis has polarized cells with nuclei located at the basal part of the cell, whereas mitochondria are in the apical one. The epidermis is entirely covered by microvilli and locomotory cilia anchored in the cytoplasm by vertical and horizontal rootlets. Rootlets exhibit distinct length and periodic structure in the two species. Furthermore, in each species vertical and horizontal rootlets possess different periodic structure. The pattern of termination of microtubules in epidermal cilia is described for the first time in the Typhloplanida; central microtubules shift along one axonemal side, doublets 1 and 6–9 lose their microtubule B, and gradually peripheral doublets become singlets. Finally, an electron-dense material caps the tip of the cilia. This pattern of termination closely resembles that of Temnocephalida, Kalytorhynchia, and Dalyelliida examined so far, but differences exist.     

The eyespots of the oncomiracidium of Euzetrema knoepffleri (Monogenea): ultrastructure and evolution during the life-cycle (author's transl)]

Euzetrema knoepffleri (Monogenea, Monopisthocotylea) is a parasite of the urinary bladder of Euproctus montanus (Amphibia, Urodela). The eyespots of the oncomiracidium of E. knoepffleri are rhabdomeric; each anterior pigmented cell has one rhabdomere, each posterior pigmented cell has two rhabdomeres. This pattern is quite similar to that of the eyespot of another Monopisthocotylea, Entobdella soleae but the lack of cristalline lens in E. knoepffleri appears as a new feature of the Monopisthocotylea. The symmetry of the pigmented structures seems to be effectively connected with the swimming mode of the larva. Moreover, the ultrastructural differences between the two species Euzetrema knoepffleri and Entobdella soleae may be in relation with their different behaviour concerning light. After the fixation of the larva on its host, the comparative study of the evolution of the eyespots, shows the disappearance of the cristalline lens in Entobdella, and the loss of rhabdomeric structures in Euzetrema. These differences seem related with the nature of the microbiotope of the adult: Entobdella soleae is a skin parasite, Euzetrema knoepffleri a reno-vesical one.     

Cytoskeletal architecture and immunocytochemical localization of fodrin in the terminal web of the ciliated epithelial cell

In order to understand the cytoskeletal architecture at the terminal web of the ciliated cell, we examined chicken tracheal epithelium by quick-freeze deep-etch (QFDE) electron microscopy combined with immunocytochemistry of fodrin. At the terminal web, the cilia ended into the basal bodies and then to the rootlets. The rootlets were composed of several filaments and globular structures attached regularly to them. Decoration with myosin subfragment 1 (S1) revealed that some actin filaments ran parallel to the apical plasma membrane between the basal bodies, and other population traveled perpendicularly or obliquely, i.e., along the rootlets. Some actin filaments were connected to the surface of the basal bodies and the basal feet. Among the basal bodies and the rootlets there existed three kinds of fine crossbridges, which were not decorated with S1. In the deeper part of the terminal web, intermediate filaments were observed between the rootlets and were sometimes crosslinked with the rootlets. Immunocytochemistry combined with the QFDE method revealed that fodrin was a component of fine crossbridges associated with the basal bodies. We concluded that an extensive crosslinker system among the basal bodies and the rootlets along with networks of actin and intermediate filaments formed a structural basis for the effective beating of cilia.     

Fine structure of the cephalic sensory organ in veliger larvae of Littorina littorea, (L.) (Mesogastropoda,Littorinidae)

The cephalic sensory organ (CSO) in planktonic veliger larvae of Littorina littorea is situated dorsally between the velar lobes at the level of the shell aperture. It consists of ciliated primary sensory cells, adjacent accessory cells and supporting epithelial cells. Cell bodies of the ciliated cells originate in the cerebral commissure and their dendrites pass to the epidermis. The flask-shaped sensory cells are characterized by a deep invaginated lumen with modified cilia arising from the cell surface in the lumen. These cilia are presumed to be non-motile because they lack striated rootlets and show a modified microtubular pattern (6 + 2, 7 + 2 and 8 + 2). The adjacent accessory cells never possess an invaginated lumen; occasionally cilia and branched microvilli arise from the apical surface. These cells may be sensory, but there is no obvious direct connection with the nervous system. The supporting epithelial cells are part of the epidermis and flank the apical necks of the sensory and accessory cells. Morphological evidence suggests that the CSO may function in chemoreception related to substrate selection at settlement, feeding or other behaviour.     

Epidermal ciliary ultrastructure of adult and larval sipunculids (Sipunculida)

The ultrastructure of the ciliary apparatus of multiciliated epidermal cells in larval and adult sipunculids is described and the phylogenetic implications discussed. The pelagosphera of Apionsoma misakianum has a dense cover of epidermal cilia on the head region. The cilia have a long, narrow distal part and two long ciliary rootlets, one rostrally and one vertically orientated. The adult Phascolion strombus has cilia on the nuchal organ and on the oral side of the tentacles. These cilia have a narrow distal part as in the A. misakianum larva, but the ciliary rootlets have a different structure. The first rootlet on the anterior face of the basal body is very short and small. The second, vertically orientated rootlet is long and relatively thick. The two ciliary rootlets present in the larval A. misakianum are similar to the basal metazoan type of ciliary apparatus of epidermal multiciliated cells and thus likely represent the plesiomorphic state. The minute first rootlet in the adult P. strombus is viewed as a consequence of a secondary reduction. No possible synapomorphic character with the phylogenetically troublesome Xenoturbella was found.     

Epidermal ultrastructure of the adult parasitic phase female and encapsulated larva of Kronborgia isopodicola (Platyhelminthes, Fecampiidae): Phylogenetic implications

The parasitic phase female K. isopodicola possesses a ciliated epidermis of polyhedral cells. Adjacent lateral plasma membranes are separated at intervals creating intercellular spaces. Epidermal cilia are anchored by a horizontal rootlet, opposite which a spur projects from the basal body, and a narrow vertical rootlet. The cytoplasm contains coated vesicles, and coated pits lie between microvilli. Large granular and vesicular bodies (rhabdoids) are scattered through the epidermal epithelium; in the epidermis of the encapsulated larva, granular rhabdoids are densely packed and slender, more compact bodies also occur. The compact, granular and vesicular bodies are probably morphological variants of the same epidermal structures, suggested to undergo sequential changes accomplished in later stages by lysosomal activity. Morphologically similar epidermal bodies are found in triclads. They are also characteristic of the parasitic genus Urastoma, which shares other ultrastructural features with K. isopodicola. The Neodermata may have arisen from parasitic turbellarian forms, at a more “primitive” level of organization than ancestors of the contemporary Rhabdocoela.     

Structure and maintenance of the epidermis in Friedmaniella sp. (Prolecithophora)

The epidermis of Friedmaniella sp. has been studied using light and electron microscopy. Three main morphological features characterize its cells, namely (1) DNA bodies in the nuclei, (2) an extensive Golgi apparatus with a well-developed system of transport vesicles, (3) clusters of centrioles mainly in the basal cytoplasm and axonemes and rootlets in the middle and apical cell parts. These peculiarities may indicate continuous physiological regeneration within the cell at the level of cell organelles. DNA bodies may prove to be a taxonomic feature distinguishing the Prolecithophora from other turbellarians.     

Ultrastructural studies of epidermis, sense receptors and sperm of Craspedella sp. and Didymorchis sp. (Platyhelminthes, Rhabdocoela)

Craspedella has a non-ciliated epidermis with nuclei located in the epidermis and with short microvilli. There is a thin basal lamina and thick underlying fibrous matrix. Rhabdites are secreted through ducts lined by microtubules. Multiciliate sense receptors consist of bundles of dendrites in a depression of the epidermis. Each dendrite has a cilium with a cross-striated rootlet; there are no electron-dense collars. Spermatozoa have peripheral microtubules which in cross-section are arranged in a ring-like or spiral fashion, numerous electron-dense granules, mitochondria and a nucleus; axonemes of the 9 +'1'type are free for most of their length. Centrioles occur in some nerve fibres. In Didymorchis parts of the epidermis are ciliated and epidermal perikarya are 'insunk', connected to the surface part of the epidermis by a single cytoplasmic process. Epidermal cilia have cross-striated vertical and horizontal rootlets. In the ciliary tips a short electron-dense rod along the central pair of tubules extends to the tip, where it widens to become a terminal plate; peripheral doublets gradually disappear by losing their microtubules. Receptors observed are uniciliate. Spermatozoa are as in Craspedella . Ultrastructural evidence indicates that Craspedella and Didymorchis arc closely related and belong to the Rhabdocoela.     

Cytoskeletal elements in arthropod sensilla and mammalian photoreceptors.

Ciliary receptor cells, typified by cilia or modified cilia, are very common in the animal kingdom. In addition to the cytoskeleton of their ciliary processes these receptors possess other specific prominent cytoskeletal elements. Two representative systems are presented: i) scolopidia, mechanosensitive sensilla of various arthropod species; and ii) photoreceptor cells of the retina of the bovine eye. Two cytoskeletal structures are characteristic for arthropod scolopidia: a scolopale typifies the innermost auxiliary cell, and long ciliary rootlets are extending well into the sensory cells. The latter element is also characteristic for the inner segment of the photoreceptor cells in bovine. The scolopale of scolopidia is mainly composed of actin filaments. In the absence of myosin, the uniform polarity of the actin filaments and their association with tropomyosin all indicate a stabilizing role of the filament bundles within the scolopale. This function and a certain elasticity of actin filament bundles may be important during stimulation of the sensilla. The ciliary rootlets of both systems originate at the basal bodies at the ciliary base of the sensory cells and project proximally. These rootlets are composed of longitudinally oriented, fine filaments forming a characteristic regular cross-striation. An alpha-actinin immunoreactivity was detected within the ciliary rootlets of scolopidia. In addition, antibodies to centrin react with the rootlets of both types of receptors. Since centrin is largely responsible for the contraction of the flagellar rootlets in green algae, contraction may also occur in the ciliary rootlets of insect sensilla and vertebrate photoreceptors. In both systems, contraction or relaxation of the ciliary rootlets could serve in sensory transduction or adaptation.     

Ultrastructure of the Flame Bulbs of Urastoma cyprinae (Platyhelminthes, ‘Prolecithophora’, Urastomidae)

The ultrastructure of the flame bulbs of the turbellarian Urastoma cyprinae from Mytilus galloprovincialis in the Mediterranean is described. The nucleus of the terminal cell is located some distance basal to the rootlets of the cilia forming the flame; the cytoplasm contains numerous tubules approximately 54–66 nm in diameter, and vesicles. Thick walled, densely packed rod-like structures coil around each other with a tendency towards longitudinal orientation close to the flame. The rod-like structures tightly surround the basal part of the flame and the distal cytoplasmic tube in the apical part of the flame. Some of them, including the inner predominantly longitudinally directed ones, are continuous with the cytoplasm of the terminal cell, others are continuous with the cytoplasm of the distal cytoplasmic tube. Internal leptotriches arise from the cytoplasm of the terminal cell and intrude between the basal parts of the cilia of the flame. The distal cytoplasmic tube possesses a septate junction. The flame bulb of Urastoma differs distinctly from those known from other Platyhelminthes; implications for the phylogeny of Platyhelminthes are discussed.     

Evolution of monogenean parasites across vertebrate hosts illuminated by the phylogenetic position of Euzetrema Combes, 1965 within the Monopisthocotylea

The Monogenea, which is divided into two clades, namely the Monopisthocotylea and Polyopisthocotylea, is a highly diversified group of platyhelminth parasites that infest mainly actinopterygian and chondrichthyan fishes but also, to a lesser extent, freshwater sarcopterygian hosts. Euzetrema knoepffleri Combes, 1965 (Monogenea: Iagotrematidae), which is specific to the salamander Euproctus montanus Savi, 1838 is among the rare monopisthocotylean parasites infesting tetrapod hosts. We sequenced the complete 18S rRNA gene of this parasite to infer its phylogenetic position within the Monopisthocotylea. Our results provide a new insight for coevolutionary scenarios between monopisthocotyleans and gnathostomatan hosts. Indeed, the basal position of E. knoepffleri within a subgroup of the Monopisthocotylea which comprises two clusters that both include parasites of the Actinopterygii and Chondrichthyes, suggests a very old association between the Iagotrematidae and tetrapods. Furthermore, if we take into account a recent view of Gnathostomata evolution where bony and cartilaginous fishes are regarded as a monophyletic group, it could be argued that the Iagotrematidae arose very early, during the fish–tetrapod transition, as did the Polystomatidae, the only monogenean family of the Polyopisthocotylea that infests sarcopterygian hosts.  © 2003 The Linnean Society of London, Biological Journal of the Linnean Society , 2003, 80 , 727–734.     

Organization of actin microfilaments in the apical border of oviduct ciliated cells

Actin microfilaments were localized in quail oviduct ciliated cells using decoration with myosin subfragment S1 and immunogold labeling. These polarized epithelial cells show a well developed cytoskeleton due to the presence of numerous cilia and microvilli at their apical pole. Most S1-decorated microfilaments extend from the microvilli downward towards the upper part of the ciliary striated rootlets with which they are connected. From the microvillous roots, a few microfilaments connect the proximal part of the basal body or the basal foot associated with the basal body. Microfilament polarity is shown by S1 arrowheads pointing away from the microvillous tip to the cell body. Furthermore, short microfilaments are attached to the plasma membrane at the anchoring sites of basal bodies and run along the basal body. The polarity of these short microfilaments is directed from the basal body anchoring fibers downward to the cytoplasm. At the cell periphery, microfilaments from microvillous roots and ciliary apparatus are connected with those of the circumferential actin belt which is associated with the apical zonula adhaerens. Together with the other cytoskeletal elements, the microfilaments increase ciliary anchorage and could be involved in the coordination of ciliary beating. Moreover, microvilli surrounding the cilia probably modify ciliary beating by offering resistance to cilium bending. The presence of microvilli could explain the fact that mainly the upper part of the cilia appanars to be involved in the axonemal bending in metazoan ciliated cells.     

THE FINE STRUCTURE OF EPENDYMA IN THE BRAIN OF THE RAT

The ciliated ependyma of the rat brain consists of a sheet of epithelial cells, the luminal surface of which is reflected over ciliary shafts and numerous evaginations of irregular dimensions. The relatively straight lateral portions of the plasmalemma of contiguous cells are fused at discrete sites to form five-layered junctions or zonulae occludentes which obliterate the intercellular space. These fusions occur usually at some distance below the free surface either independently or in continuity with a second intercellular junction, the zonula adhaerens. The luminal junction is usually formed by a zonula adhaerens or, occasionally, by a zonula occludens. The finely granular and filamentous cytoplasm contains supranuclear dense bodies, some of which are probably lysosomes and dense whorls of perinuclear filaments which send fascicles toward the lateral plasmalemma. The apical regions of the cytoplasm contain the basal body complexes of neighboring cilia. These complexes include a striated basal foot and short, non-striated rootlets emanating from the wall of each basal body. The rootlets end in a zone of granules about the proximal region of the basal body, adjacent to which may lie a striated mass of variable shape. All components of the basal body complex of adjacent cilia are independent of each other.     

Morphology and ultrastructure of the sensory spine,a presumed mechanoreceptor of Sphaeroma hookeri (Crustacea,Isopoda), and remarks on similar spines in other peracarids

The isopod Sphaeroma hookeri and many other isopods and peracarids have a sensory spine with laterally inserting sensory hair, positioned in the apical region of the propodal palm of pereopod 1. This spine is innervated by five to eight sensory cells (each giving rise to one cilium) the dendrites of which can be divided into an inner and outer dendritic segment. The cilia are surrounded by an extracellular, electron-dense dendritic sheath. Thirteen enveloping cells are present. The outer dendritic segment (structure beyond the basal bodies) contains two receptor lymph cavities; the inner one lying within the dendritic sheath is homologous with the inner receptor lymph cavity of insects. Scolopales, or tubular bodies, are lacking; their function is probably accomplished by the dendritic sheath. Apically the sensory hair does not have a pore, and the spine is heavily sclerotized. The inner dendritic segment begins with a basal body from which rootlets of different length and thickness extend into the dendrite. In the latter is an accumulation of vesicles. The dendrites keep close contact with other dendrites and the enveloping cells by desmosomal membrane structures. The possible importance of the sensory spine for phylogenetic studies is discussed.     

The absolute configuration of the flagellar apparatus in zoospores from two species ofLaminariales (Phaeophyceae

Summary We examined the zoospores produced by the unilocular sporangia ofLaminaria digitata (L.) Lamour. andNereocystis luetkeana Post. & Rupr. by serial sectioning to determine the absolute configuration of their flagellar apparatuses. The basal bodies, which are interconnected by three striated bands, lie parallel to the ventral face of the zoospore, and the posterior basal body always is found to the right of the anterior basal body when the cell is viewed from the ventral face, anterior end up. The four rootlets associated with the basal bodies include a major anterior rootlet of about seven microtubules extending from the anterior basal body along the ventral face towards the apex, a five-membered bypassing rootlet that passes ventral to the basal bodies and is connected to the posterior basal body by a posterior fibrous band, and two short rootlets having a single member each, the minor anterior and posterior rootlets. We consider the configuration observed here to be typical of most phaeophycean motile cells. The flagellar apparatus features suggest a considerable phylogenetic difference between thePhaeophyceae and other classes of chlorophyll c-containing organisms.     

The ciliary basal apparatus is adapted to the structure and mechanics of the epithelium

The basal apparatuses which anchor the gill cilia in Branchiostoma lanceolatum (Pallas) and the actinopharynx cilia in Calliactis parasitica (Couch) are similar in structure. In C. parasitica the pharynx epithelium and the basal apparatuses are flexible. The basal apparatuses, however, bend in only one direction. This mechanism may permit epithelial flexibility whilst maintaining a similar basal orientation between cilia. In B. lanceolatum the ciliated gill epithelia are mechanically stable but the epithelial surfaces are curved. The basal apparatuses may correct for this curvature, with short rootlets between the distal centrioles (basal bodies) and the cell membranes, so that their cilia also share a common orientation. A common basal orientation between cilia is important for their coordination. The degree of coordination depends upon the function of the cilia; water-propelling cilia are more precisely coordinated than mucus-propelling cilia. Much of the structural diversity of ciliary basal apparatuses in Metazoa may be due to variation in the demands of anchoring functionally different cilia to epithelia which have different structural and mechanical properties.     

ULTRASTRUCTURE OF THE FLAGELLAR APPARATUS OF PYROBOTRYS (CHLOROPHYCEAE)1

The flagellar apparatus of Pyrobotrys has a number of features that are typical of the Chlorophyceae, but others that are unusual for this class. The two flagella are inserted at the apex, but they extend to the side of the cell toward the outside of the colony, here designated as the ventral side. Four basal bodies are present, two of which extend into flagella. Four microtubular rootlets alternate between the functional and accessory basal bodies. In each cell, the two ventral rootlets are nearly parallel, but the dorsal rootlets are more widely divergent. The rootlets alternate between two and four microtubules each. A striated distal fiber connects the two functional basal bodies in the plane of the flagella. Two additional, apparently nonstriated, fibers connect the basal bodies proximal to the distal fiber. Another striated fiber is associated with each four-membered rootlet near its insertion into the flagellar apparatus. A fine periodic component is associated with each two-membered rootlet. A rhizoplast-like structure extends into the cell from each of the functional basal bodies. The arrangement of these components does not reflect the 180° rotational symmetry that is usually present in the Chlorophyceae, but appears to be derived from a more symmetrical ancestor. It is suggested that the form of the flagellar apparatus is associated with the unusual colony structure of Pyrobotrys.