Nervous system and musculature of the parasitic turbellarian Notentera ivanovi (Plathelminthes, Fecampiida).

The article reports the first description of the architecture of the musculature and cholinergic nervous system of the parasitic turbellarian Notentera ivanovi from the White Sea using histochemistry and confocal scanning laser microscopy. It has been demonstrated that the body wall is composed of layers of circular and longitudinal muscles between which there are diagonal muscles described here for the first time. The nervous system is of the regular closely spaced orthogon type with pronounced radiality. In order to make it clear whether the orthogon of this type is a phylogenetic characteristic of this group of worms or its shape is associated with the flat rounded body, the study of other representatives of the Fecampiida group is required.

Muscular system of a peculiar parasitic cnidarian Polypodium hydriforme: a phalloidin fluorescence study.

Musculature of the free-living stages of Polypodium hydriforme has been studied using phalloidin fluorescence method and confocal microscopy. P. hydriforme is a unique cnidarian possessing only smooth muscle cells situated within the mesoglea, not epithelial muscle cells, like the rest of cnidarians. Phalloidin fluorescence on whole mount preparations demonstrates an extensively developed subepidermal muscle system mostly consisting of long parallel fibers running along the tentacles. For the first time along with contracted muscle fibers we could clearly demonstrate relaxed fibers looking as long spirals. System of thin parallel circular F-actin positive fibers has been discovered outside of longitudinal muscles. The body of the animal and the mouth cone contain weakly developed parallel muscles. No special attachment of the muscle fibers to the tips of the tentacles or to the rim of the mouth has been observed. The results are discussed in connection with the "triploblastic" organization of P. hydriforme and its phylogenetic position.

Copulatory organ musculature in Childia (Acoela) as revealed by phalloidin fluorescence and confocal microscopy.

Copulatory organs of eight species of the monophyletic taxon Childia were investigated in detail, using phalloidin fluorescence method and confocal microscopy. Childia species were shown to have one, two or several tubular stylets, conical to cylindrical in shape, composed of few to numerous needles. The musculature varied greatly, from the absence of seminal vesicle to extensively developed seminal vesicles with several additional types of specialized muscles. Ten copulatory organ characters were coded and mapped on the total evidence tree. The data obtained permitted to follow the evolution of the Childia stylet and to demonstrate that the structure of the stylet apparatus is largely consistent with the phylogeny of the group (CI=0.75). Possible function of different muscle specializations was discussed.

F-actin and beta-tubulin localization in the myxospore stinging apparatus of Myxobolus pseudodispar Gorbunova, 1936 (Myxozoa, Myxosporea).

To understand the discharge mechanism of Myxozoan polar capsule (cnida) it is necessary to verify the role of major cytoskeletal proteins in the process. With this aim F-actin and beta-tubulin localization in spores of myxosporean developmental phase (in myxospores) of Myxobolus pseudodispar Gorbunova, 1936 has been studied under confocal scanning laser microscope using phalloidin fluorescent staining of F-actin and indirect anti-beta-tubulin immunostaining. F-actin has been detected in walls of the stinging tube invaginated into the polar capsule of myxospore. The fact suggests the contractile proteins involvement in the process of myxozoan polar capsule extrusion. In addition, the cytoplasm of amoeboid sporoplasm inside the spore cavity is stained by phalloidin. A polar cap with strong beta-tubulin immunoreacton is observed at the front pole of fully mature myxospore above the outlets of the polar capsule discharge channels. The role of the beta-tubulin cap is supposed to be similar to that of the cnidarian cnidocil made of microtubules. The weaker beta-tubulin immunoreactivity has been found in stinging tubes, in polar capsule walls as well as in the suture line of spore walls and in the cytoplasm of amoeboid sporoplasm. The involvement of cytoskeletal proteins in the process of polar capsule extrusion is discussed. A hypothesis on the myxozoan polar capsule discharge mechanism is suggested. The mechanism of myxozoan cnida discharge is compared with that of cnidaria.

Musculature of an illoricate predatory rotifer Asplanchnopus multiceps as revealed by phalloidin fluorescence and confocal microscopy.

The pattern of muscles in the actively swimming predatory rotifer Asplanchnopus multiceps is revealed by staining with tetramethyl-rhodamine isothiocyanate (TRITC)-labelled phalloidin and confocal scanning laser microscopy (CSLM). The major components of the musculature are: prominent semicircular muscles of the corona; paired lateral, dorsal and ventral retractors in the trunk; a network of six seemingly complete circular muscles and anastomosing longitudinal muscles in the trunk; two short foot retractors, originating from a transverse muscle in the lower third of the trunk. The sphincter of the corona marks the boundary between the head and the trunk. The muscular patterns in rotifers with different lifestyles differ clearly, therefore, the muscular patterns seem to be determined by the mode of locomotion and feeding behaviour.

Neuropeptides in flatworms.

The use of well-characterized antibodies raised to neuronal signal substances and their application through immunocytochemistry and confocal scanning laser microscopy has revolutionized studies of the flatworm nervous system (NS). Data about flatworm neuropeptides and the spatial relationship between neuropeptides and other neuronal signal substances and muscle fibers are presented. Neuropeptides form a large part of the flatworm NS. Neuropeptides are especially important as myoexcitatory transmitters or modulators, controlling the musculature of the attachment organs, the stomatogastric and the reproductive systems.

The nervous and muscular systems in the free-living flatworm Castrella truncata (Rhabdocoela): an immunocytochemical and phalloidin fluorescence study.

In order to broaden the information about the organisation of the nervous system in the Plathelminthes, an immunocytochemical (ICC) study of the free-living flatworm Castrella truncata (Dalyellioida) has been performed. This is the first time a representative of the taxon Rhabdocoela has been studied with the ICC technique. Antibodies to 5-HT and FMRF-amide and confocal scanning laser microscope were used. 5-HT and FMRF-amide immunoreactivity was observed in the neuropile of the brain, in the cerebral neurones, in the three pairs of longitudinal nerve cords and the adjoining neurones, in the pharyngeal nerve ring and in twelve neurones associated with the pharynx. 5-HT and FMRF-amide immunoreactivity occurs in separate sets of neurones. Only FMRF-amide immunoreactivity was observed in a peculiar ellipse-shaped structure in the brain, and in the genital plexus. The type of orthogon is discussed. Staining of the muscular system with TRITC conjugated phalloidin revealed muscle patterns that have not been described previously.

Organisation of the nervous system in the Acoela: an immunocytochemical study.

In order to broaden the information about the organisation of the nervous system in taxon Acoela, an immunocytochemical study of an undetermined Acoela from Cape Kartesh, Faerlea glomerata, Avagina incola and Paraphanostoma crassum has been performed. Antibodies to 5-HT and the native flatworm neuropeptide GYIRFamide were used. As in earlier studies, the pattern of 5-HT immunoreactivity revealed an anterior structure composed mainly of commissures, a so-called commissural brain. Three types of brain shapes were observed. No regular orthogon was visualised. GYIRFamide immunoreactive cell clusters were observed peripherally to the 5-HT immunoreactive commissural brain. Staining with anti-GYIRFamide revealed more nerve processes than did staining with anti-FMRFamide. As no synapomorphies were found in the organisation of the nervous system of the Acoela and that of the Platyhelminthes, the results support the view that the Acoela is not a member of the Platyhelminthes.

The brain of the Nemertodermatida (Platyhelminthes) as revealed by anti-5HT and anti-FMRFamide immunostainings.

The taxa Nemertodermatida and Acoela have traditionally been considered closely related and classified as sister groups within the Acoelomorpha Ehlers 1984 (Platyhelminthes). Recent molecular investigations have questioned their respective position. In this study, the 5-HT and FMRFamide immunoreactivity (IR) in the nervous system of two nemertodermatids, Nemertoderma westbladi and Meara stichopi, is described. The 5-HT immunoreactive pattern differs in the two nemertodermatids studied. In M. stichopi, two loose longitudinal bundles of 5-HT-immunoreactive fibres and an basi-epidermal nerve net were observed. In N. westbladi the 5-HT-IR shows a ring-shaped commissural structure, different from the commissural brain of acoels. In both nemertodermatids, FMRFamide immunoreactive nerve fibres followed the 5-HT-immunoreactive fibres. It is demonstrated that the Nemertodermatida have neither a 'commissural brain' structure similar to that of the Acoela, nor a 'true', ganglionic brain and orthogon, typical for other Platyhelminthes. The question of the plesiomorphic or apomorphic nature of the nervous system in Nemertodermatida cannot yet be answered. The neuroanatomy of the studied worms provides no synapomorphy supporting the taxon Acoelomorpha.

An endocrine brain? The pattern of FMRF-amide immunoreactivity in Acoela (Plathelminthes).

In order to solve the problem of whether the Acoela have a brain of a common flatworm type, an immunocytochemical study was performed of the pattern of FMRF-amide in the nervous system of four species of Acoela, Anaperus biaculeatus, Childia groenlandica, Antinoposthia beklemischevi and Mecynostomum sp. In all species a FMRF-amide positive bilobed brain-like structure lacking neuropile was observed. This brain is composed of large multipolar nerve cells with short processes. The FMRF-amide immunoreactivity appears as spots in the cytoplasm. Short lateral branches originating from the periphery of the brain were observed in Anaperus biaculeatus. No immunoreactive nerve cords were detected. The results indicate that the brain-like structure of Acoela is not homologous with the brains of other Plathelminthes and implies an independent evolution of Acoela.