Ciliary Tufts Research Articles

The ultrastructure of the apical organ of Curini-Galletti's larva, a new polyclad larval type.

Polycladida are the only free-living flatworms with a planktonic larval stage in some species. Currently, it is not clear if a larval stage is ancestral in polyclads, and which type of larva that would be. Known polyclad larvae are Müller's larva, Kato's larva and Goette's larva, differing by body shape and the number of lobes and eyes. A valuable character for the comparison and characterisation of polyclad larval types is the ultrastructural composition of the apical organ. This organ is situated at the anterior pole of the larva and is associated with at least one ciliary tuft. The larval apical organ of Theama mediterranea features two multiciliated apical tuft sensory cells. Six unfurcated apical tuft gland cell necks are sandwiched between the apical tuft sensorycells and two anchor cells and have their cell bodies located lateral to the brain. Another type of apical gland cell necks is embedded in the anchor cells. Ventral to the apical tuft, ciliated sensory neurons are present, which are neighbouring the cell necks of two furcated apical tuft glandcells. Based on the ultrastructural organisation of the apical organ and other morphological features, like a laterally flattened wedge-shaped body and three very small lobes, we recognise the larva of T. mediterranea as a new larval type, which we name Curini-Galletti's larva after its first discoverer. The ultrastructural similarities of the apical organ in different polyclad larvae support their possible homology, that is,all polyclad larvae have likely evolved from a common larva.

Gene Regulatory Network that Shaped the Evolution of Larval Apical Organ in Cnidaria.

Among non-bilaterian animals, a larval apical sensory organ with integrated neurons is only found in cnidarians. Within cnidarians, an apical organ with a ciliary tuft is mainly found in Actiniaria. Whether this apical tuft has evolved independently in Actiniaria or alternatively originated in the common ancestor of Cnidaria and Bilateria and was lost in specific groups is uncertain. To test this hypothesis, we generated transcriptomes of the apical domain during the planula stage of four species representing three key groups of cnidarians: Aurelia aurita (Scyphozoa), Nematostella vectensis (Actiniaria), and Acropora millepora and Acropora tenuis (Scleractinia). We showed that the canonical genes implicated in patterning the apical domain of N. vectensis are largely absent in A. aurita. In contrast, the apical domain of the scleractinian planula shares gene expression pattern with N. vectensis. By comparing the larval single-cell transcriptomes, we revealed the apical organ cell type of Scleractinia and confirmed its homology to Actiniaria. However, Fgfa2, a vital regulator of the regionalization of the N. vectensis apical organ, is absent in the scleractinian genome. Likewise, we found that FoxJ1 and 245 genes associated with cilia are exclusively expressed in the N. vectensis apical domain, which is in line with the presence of ciliary apical tuft in Actiniaria and its absence in Scleractinia and Scyphozoa. Our findings suggest that the common ancestor of cnidarians lacked a ciliary apical tuft, and it could have evolved independently in the Actiniaria.

Integrative taxonomy and molecular phylogeny of three poorly known tintinnine ciliates, with the establishment of a new genus (Protista; Ciliophora; Oligotrichea)

BackgroundThe taxonomic classification of the suborder Tintinnina Kofoid & Campbell, 1929, a species-rich group of planktonic ciliated protistans with a characteristic lorica, has long been ambiguous largely due to the lack of cytological and molecular data for most species. Tintinnopsis is the largest, most widespread, and most taxonomically complex genus within this group with about 170 species occurring in nearshore waters. Previous molecular phylogenetic studies have revealed that Tintinnopsis is polyphyletic.ResultsHere we document the live morphology, infraciliature, gene sequences, and habitat characteristics of three poorly known tintinnine species, viz. Tintinnopsis karajacensis Brandt, 1896, Tintinnopsis gracilis Kofoid & Campbell, 1929, and Tintinnopsis tocantinensis Kofoid & Campbell, 1929, isolated from the coastal waters of China. Based on a unique cytological feature (i.e., an elongated ciliary tuft with densely arranged kinetids) in the former two species, Antetintinnopsis gen. nov. is erected with Antetintinnopsis hemispiralis (Yin, 1956) comb. nov. (original combination: Tintinnopsis hemispiralis Yin, 1956) designated as the type species. Moreover, A. karajacensis (Brandt, 1896) comb. nov. (original combination: Tintinnopsis karajacensis Brandt, 1896) and A. gracilis (Kofoid & Campbell, 1929) comb. nov. (original combination: Tintinnopsis gracilis Kofoid & Campbell, 1929) are placed in a highly supported clade that branches separately from Tintinnopsis clades in phylogenetic trees based on SSU rDNA and LSU rDNA sequence data, thus supporting the establishment of the new genus. One other species is assigned to Antetintinnopsis gen. nov., namely A. subacuta (Jörgensen, 1899) comb. nov. (original combination Tintinnopsis subacuta Jörgensen, 1899).ConclusionsThe findings of the phylogenetic analyses support the assertion that cytological characters are taxonomically informative for tintinnines. This study also contributes to the broadening of our understanding of the tintinnine biodiversity and evolution.

Morphology and Molecular Phylogeny of Four Trachelocercid Ciliates (Protozoa, Ciliophora, Karyorelictea) Found in Marine Coastal Habitats of Northern China, With Description of a New Genus, Two New Species and a New Combination

The morphology of four trachelocercid ciliates, Paratrachelocerca typica gen. nov., spec. nov., Trachelolophos monocaryon (Dragesco, 1965) comb. nov. (original combination: Tracheloraphis monocaryonDragesco, 1965), Tracheloraphis katzae spec. nov., and Tracheloraphis colubis (Kahl, 1933) Xu et al., 2011 were studied in live and protargol-stained specimens. All samples were isolated from the intertidal zone of sandy beaches at Qingdao, China. The new genus Paratrachelocerca can be distinguished from other trachelocercid genera mainly by the three circumoral kineties each composed of a row of dikinetids and the absence of a brosse or ciliary tuft in the oral cavity. The detailed investigation on the poorly described Tracheloraphis monocaryon (Dragesco, 1965) reveals that its oral infraciliature includes one uninterrupted circumoral kinety and a conspicuous ciliary tuft in the center of the oral cavity, which is consistent with the genus Trachelolophos rather than Tracheloraphis. Therefore, this species is transferred to Trachelolophos as Trachelolophos monocaryon (Dragesco, 1965) comb. nov. Tracheloraphis katzae spec. nov. can be recognized by the combination of its minute brownish cortical granules and 9–15 somatic kineties. The small subunit (SSU) rDNA of each species was sequenced for the first time. Phylogenetic analyses of the SSU rDNA show that Paratrachelocerca typica gen. nov., spec. nov. clusters with Apotrachelocerca arenicola (Kahl, 1933) Xu et al., 2011 in a group that is sister to all other trachelocercids.

Three redescriptions in Tintinnopsis (Protista: Ciliophora: Tintinnina) from coastal waters of China, with cytology and phylogenetic analyses based on ribosomal RNA genes

BackgroundThe taxonomy of tintinnine ciliates is vastly unresolved because it has traditionally been based on the lorica (a secreted shell) and it has only recently incorporated cytological and molecular information. Tintinnopsis, the most speciose tintinnine genus, is also the most problematic: it is known to be non-monophyletic, but it cannot be revised until more of its species are studied with modern methods.ResultsHere, T. hemispiralis Yin, 1956, T. kiaochowensis Yin, 1956, and T. uruguayensis Balech, 1948, from coastal waters of China, were studied. Lorica and cell features were morphometrically investigated in living and protargol-stained specimens, and sequences of three ribosomal RNA (rRNA) loci were phylogenetically analyzed. The three species show a complex ciliary pattern (with ventral, dorsal, and posterior kineties and right, left, and lateral ciliary fields), but differ in lorica morphology, details of the somatic ciliature and rRNA gene sequences. Tintinnopsis hemispiralis is further distinguished by a ciliary tuft (a ribbon of very long cilia originated from the middle portion of the ventral kinety and extending out of the lorica) and multiple macronuclear nodules. Both T. kiaochowensis and T. uruguayensis have two macronuclear nodules, but differ in the number of somatic kineties and the position of the posterior kinety. Two neotypes are fixed for T. hemispiralis and T. kiaochowensis to stabilize the species names objectively, mainly because of the previous unavailability of type materials. By phylogenetic analysis and comparison with closely-related species, we infer that the ciliary tuft and details such as the commencement of the rightmost kinety in the lateral ciliary field are synapomorphies that may help clarify the systematics of Tintinnopsis-like taxa.ConclusionThe redescriptions of three poorly known Tintinnopsis species, namely T. hemispiralis, T. kiaochowensis, and T. uruguayensis firstly revealed their ciliary patterns and rRNA sequences. This study expands knowledge and database of tintinnines and helps in identifying potential synapomorphies for future taxonomic rearrangements.

Morpholino-Mediated Knockdown of Ciliary Genes in Euplotes vannus, a Novel Marine Ciliated Model Organism.

Cilia are highly conserved organelles present in almost all types of eukaryotic cells, and defects in cilia structure and/or function are related to many human genetic disorders. Single-celled ciliated protists, which possess diverse types of cilia, are remarkable model organisms for studying cilia structures and functions. Euplotes vannus is a representative ciliate with many intriguing features; for example, it possesses extensively fragmented somatic genomes and a high frequency of + 1 programmed ribosomal frameshifting. However, the molecular mechanisms underlying these remarkable traits remain largely unknown, mainly due to the lack of efficient genetic manipulation tools. Here, we describe the first application of a morpholino-based strategy to knockdown gene expression in E. vannus. Through interfering with the function of two ciliary genes, ZMYND10 and C21ORF59, we showed that these two genes are essential for the ciliary motility and proliferation of E. vannus cells. Strikingly, both ZMYND10- and C21ORF59-knockdown cells developed shorter cilia in the ventral cirri, a special type of ciliary tuft, suggesting a novel role for these genes in the regulation of cilia length. Our data provide a new method to explore gene function in E. vannus, which may help us to understand the functions of evolutionarily conserved cilia-related genes as well as other biological processes in this intriguing model.

Release of Sticky Glycoproteins from Chlamydomonas Flagella During Microsphere Translocation on the Surface Membrane.

Chlamydomonas flagella display surface motility such that small polystyrene beads (microspheres) attached to the flagellar membrane move bidirectionally along the flagellum. This surface motility enables cells to glide on a solid substrate to which they are attached by the flagellar surface. Previous studies suggested that microsphere movement and gliding motility result from the movement of transmembrane glycoprotein(s) within the plane of the plasma membrane, driven by intraflagellar transport (IFT), which utilizes cytoplasmic dynein and kinesin-2. However, it is not well understood how a cell can continuously glide in one direction further than a single flagellar length. Here we show that, during microsphere translocation on the flagella of a non-motile mutant, pf18, some flagellar glycoproteins, including FMG-1B and FAP113, detach from the membrane and attach to the microspheres. We propose that such relocation of surface glycoproteins underlies the ability to glide over a long distance. Surface motility is likely common to cilia/flagella of various organisms, as a similar microsphere movement is observed in the apical ciliary tuft in sea urchin embryos.

Two new meiofaunal species of Trilobodrilus (Dinophilidae, Annelida) from California, USA

We describe two new species of the annelid genus Trilobodrilus Remane, 1925 (Dinophilidae Verill, 1892) from an intertidal and a subtidal location in San Diego, California. These two species show morphological and molecular divergences between each other and the previously described, geographically distant species. Intertidal T. windansea sp. nov. differs from subtidal T. ellenscrippsae sp. nov. most remarkably in the number and pattern of ciliary tufts and bands on the prostomium and along the body length, besides showing ca 15% difference in gene fragments of COI and CytB. Trilobodrilus windansea sp. nov., though nesting with T. ellenscrippsae sp. nov. in the molecular phylogenetic analyses, morphologically resembles the Japanese T. itoi Kajihara, Ikoma, Yamasaki & Hiruta, 2015 most closely, but still differs from this species in the higher number of apical ciliary tufts, an additional ciliary row posterior to the second ciliary band, and by lacking a forth ciliary band and segmentally arranged lateral ciliary tufts. Trilobodrilus ellenscrippsae sp. nov. is morphologically most similar to the Japanese T. nipponicus Uchida & Okuda, 1943, but is much shorter, has more apical ciliary tufts, and less regularly arranged lateral ciliary tufts along the body. All species differ significantly in all compared gene fragments, and no obvious correlation was found between habitat and the species morphology or relationships.

Prototapirella ciliates from wild habituated Virunga mountain gorillas (Gorilla beringei beringei) in Rwanda with the descriptions of two new species

The morphology of Prototapirella fosseyi n. sp., P. rwanda n. sp. and P. gorillaeImai, Ikeda, Collet, and Bonhomme, 1991 in the Entodiniomorphida were described from the mountain gorillas, Gorilla beringei beringei, in Rwanda. The ciliates have a retractable adoral ciliary zone, four non-retractable ciliary tufts in four caudalia, and one broad skeletal plate beneath the body surface. P. rwanda has a dorsal lobe and ventral lobes in two rows whereas P. fosseyi has no lobes. These two new species have an elongated body, a flat tail flap leaning to the ventral, a macronucleus with a tapering anterior end, a round posterior end and a shallow depression on the dorsal side, a micronucleus lying near the anterior end of macronucleus, a thin left region of the skeletal plate, a distinct skeletal rod plate, and four contractile vacuoles. P. gorillae has some variations in the nuclei and the skeletal plate. The infraciliary bands of three Prototapirella species were the same as some Triplumaria species; a C-shaped adoral polybrachykinety, a slender perivestibular polybrachykinety, and paralabial kineties in their retractable adoral ciliary zone and short lateral polybrachykineties in their four caudalia. The perivestibular polybrachykinety is joined only to the right end of adoral polybrachykinety.

Morphological analysis of the vestibular system of guinea pigs poisoned by organophosphate

IntroductionThe vestibular system is responsible for body balance. There are substances that damage it, causing dizziness; these are termed vestibulotoxic substances. Agrochemicals have been investigated for ototoxicity because of studies that identified dizziness as a recurrent symptom among rural workers’ complaints. ObjectiveTo histopathologically evaluate the vestibular system in guinea pigs exposed to an organophosphate, and to identify the drug's effects on this system. MethodsExperimental clinical study. Eighteen guinea pigs were used; six of them poisoned with the organophosphate chlorpyrifos at doses of 0.5mg/kg/day and seven of them at 1mg/kg/day; and a control group of five guinea pigs was exposed to distilled water, all for 10 consecutive days. Later, ciliary tufts of saccule and utricle maculae were counted by scanning electron microscopy. ResultsComparing the groups, a one-way ANOVA test for the variable “saccule” (p=0.0569) and a Kruskal–Wallis test for the variable “utricle” (p=0.8958) were performed, revealing no difference among groups in both variables. ConclusionThe histopathologic analysis of the vestibular system of guinea pigs exposed to an organophosphate showed no difference in the amount of ciliary tufts of saccule and utricle maculae at the doses tested, although the result for the variable “saccule” was considered borderline, showing a trend for significance.

Detached ciliary tufts masquerading as free-living amoebae

Detached ciliary tufts masquerading as free-living amoebae

Oestradiol and progesterone differentially alter cytoskeletal protein expression and flame cell morphology in Taenia crassiceps

We examined the effects of oestradiol (E2) and progesterone (P4) on cytoskeletal protein expression in the helminth Taenia crassiceps — specifically actin, tubulin and myosin. These proteins assemble into flame cells, which constitute the parasite excretory system. Total protein extracts were obtained from E2- and P4-treated T. crassiceps cysticerci and untreated controls, and analysed by one- and two-dimensional protein electrophoresis, flow cytometry, immunofluorescence and videomicroscopy. Exposure of T. crassiceps cysticerci to E2 and P4 induced differential protein expression patterns compared with untreated controls. Changes in actin, tubulin and myosin expression were confirmed by flow cytometry of parasite cells and immunofluorescence. In addition, parasite morphology was altered in response to E2 and P4 versus controls. Flame cells were primarily affected at the level of the ciliary tuft, in association with the changes in actin, tubulin and myosin. We conclude that oestradiol and progesterone act directly on T. crassiceps cysticerci, altering actin, tubulin and myosin expression and thus affecting the assembly and function of flame cells. Our results increase our understanding of several aspects of the molecular crosstalk between host and parasite, which might be useful in designing anthelmintic drugs that exclusively impair parasitic proteins which mediate cell signaling and pathogenic reproduction and establishment.

The diversity, development and evolution of polyclad flatworm larvae

Polyclad flatworms offer an excellent system with which to explore the evolution of larval structures and the ecological and developmental mechanisms driving flatworm and marine invertebrate life history evolution. Although the most common mode of development in polyclads might be direct development (where the embryo develops directly into a form resembling the young adult), there are many species that develop indirectly, through a planktonic phase with transient larval features, before settling to the sea floor. In this review, I introduce polyclad life history strategies, larval diversity and larval anatomical features (presenting previously unpublished micrographs of a diversity of polyclad larvae). I summarize what is known about polyclad larval development during the planktonic phase and the transition to the benthic juvenile. Finally, I discuss evolutionary and developmental scenarios on the origin of polyclad larval characters.The most prominent characters that are found exclusively in the larval stages are lobes that protrude from the body and a ciliary band, or ciliary tufts, at the peripheral margins of the lobes. Larvae with 4–8 and 10 lobes have been described, with most indirect developing species hatching with 8 lobes. A ventral sucker develops in late stage larvae, and I put forward the hypothesis that this is an organ for larval settlement for species belonging to the Cotylea. Historically, the biphasic life cycle of polyclads was thought to be a shared primitive feature of marine invertebrates, with similarities in larval features among phyla resulting from evolutionary conservation. However, our current understanding of animal phylogeny suggests that indirect development in polyclads has evolved independently of similar life cycles found in parasitic flatworms and some other spiralian taxa, and that morphological similarities between the larvae of polyclads and other spiralians are likely a result of convergent evolution.

Microanatomy and ultrastructure of outer mantle epidermis of the cuttlefish, Sepia esculenta (Cephalopoda: Sepiidae)

This study describes the ultrastructural characteristics of external epidermis of mantle of Sepia esculenta using light and electron microscopy. The epidermis was thicker on the ventral surface than on the dorsal surface, with a higher secretory cell distribution on the ventral surface than on the dorsal surface. The epidermis was a single layer composed of epithelial cells, secretory cells, ciliated cells and neuroglial cells. Epithelial cells were columnar with well-developed microvilli on the free surface, and the microvilli were covered with glycocalyx. The epithelial cells were connected to the neighboring cells by tight junctions and membrane interdigitations of the apico-frontal surface. Well-developed microfilaments were arranged in a vertical direction in the cortical cytoplasm. The secretory cells were categorized into three types (A, B and C) in accordance with the light microscopical characteristics and ultrastructures of the secretory granules. The distribution of these cells was in the following order: Type A>Type B>Type C. SEM observation revealed that the secretory pore size of the Type A secretory cells was approximately 8.6μm×12.2μm. Cytoplasm displayed a red color as the result of Masson's trichrome stain and H–E stain, and contained polygonal granules of approximately 1.2μm2 with a high electron density. The secretory pore size of the Type B secretory cells was approximately 10.1μm×12.1μm. As the results of AB–PAS (pH 2.5) and AF–AB (pH 2.5) reactions, the cytoplasm displayed a red color. The cells contained membrane bounded secretory granules with very low electron density. The secretory pore of the Type C secretory cells was circular shape, and approximately 5.5μm×5.5μm. Cytoplasm was found to be homogeneous under H–E stain and Masson's trichrome stain, and displayed a red color. As the result of AB–PAS (pH 2.5) reaction, the cytoplasm displayed a red color. The electron density of the secretory substance was the highest among the three types of secretory cells. The ciliated cells had a ciliary tuft on the free surface and were distributed throughout the mantle with the exception of the adhesive organs. Neuroglial cells were connected to the basal membrane, epithelial cells, secretory cells and nerve fibers through cytoplasmic process, and contained neurosecretory granules with high electron density within the cytoplasm.

Immunohistochemical and ultrastructural studies on ciliary sense organs of arrow worms (Chaetognatha)

We examined epibenthic and pelagic species of Chaetognatha (Spadellidae and Sagittidae) using immunohistofluorescence and confocal laser scanning microscopy to detect tubulin and cell nuclei in whole-mount preparations and scanning and transmission electron microscopy to visualize the ultrastructural organisation of their ciliary sense organs. All chaetognaths bear three types of ciliary sense organs distributed throughout the body: (1) transversally oriented ciliary fence organs, (2) longitudinally (parallel to the anterior-posterior axis) oriented ciliary tuft organs, and (3) a ciliary loop, the corona ciliata. This study targets the ciliary fence as well as the ciliary tuft organs. Two types of primary receptor cells constitute the ciliary fence and ciliary tuft organs. The first type of receptor cells forms a single cell line along the midline axis of the organs, whereas the second type of receptor cells forms multiple lines of cells at either side of type 1 cells. Each receptor cell extends a single, non-locomotory cilium from its narrow apex collared by slender, non-reinforced microvilli; however, both types of sensory cells considerably differ on ultrastructural level. Type 1 sensory cells have thicker cilia than those protruded by the type 2 sensory cells which are characterized by rootlets consisting of an elongated, amorphous distal as well as a cross-striated proximal portion. These results likely reveal that both types of sensory cells have distinct functions.

Glutathione transferase theta in apical ciliary tuft regulates mechanical reception and swimming behavior of Sea Urchin Embryos

An apical tuft, which is observed in a wide range of embryos/larvae of marine invertebrates, is composed of a group of cilia that are longer and less motile than the abundant lateral cilia covering the rest of the embryonic surface. Although the apical tuft has been thought to function as a sensory organ, its molecular composition and roles are poorly understood. Here, we identified a glutathione transferase theta (GSTT) as an abundant and specific component of the apical tuft in sea urchin embryos. The expression of GSTT mRNA increases and becomes limited to the animal plate of the mesenchyme blastula, gastrula, and prism larva. Electron microscopy and tandem mass spectrometry demonstrated that the apical tuft contains almost every axonemal component for ciliary motility. Low concentrations of an inhibitor of glutathione transferase bromosulphophthalein (BSP) induce bending of apical tuft, suggesting that GSTT regulates motility of apical tuft cilia. Embryos treated with BSP swim with normal velocity and trajectories but show less efficiency of changing direction when they collide with an object. These results suggest that GSTT in the apical tuft plays an important role in the mechanical reception for the motility regulation of lateral motile cilia in sea urchin embryos.

Light and Scanning Electron Microscope Examination of the Digestive Tract in Peppered Moray Eel, Gymnothorax pictus (Elopomorpha)

The morphology of the digestive tract of the peppered moray eel, Gymnothorax pictus (G. pictus) (Elopomorpha: Anguilliformes) was examined using both light and scanning electron microscopy. The digestive tract is composed of the esophagus, the stomach, and the intestines; pyloric caeca were absent. The stomach was divided into a cardiac region that was continuous with the esophagus, a body which terminated in a long blind sac, and a pyloric region that was continuous with the intestine. The short intestine possessed several partitions that were created by the mucosal folds within the posterior region. The terminal region of the stomach was characterized by the thick longitudinal muscularis and subserosa, and the gastric glands and microvilli were absent. Ciliary tufts of ciliated cells were observed on the surface of the partition-like mucosal folds within the intestinal wall. Acidic mucus was secreted throughout the digestive tract. It was suggested that the terminal region of the stomach is specialized for storage of large food items. In addition, it is possible that the partition-like mucosal folds within the intestine perform a function similar to that of the spiral valve and, and along with ciliated cells, facilitated digestion and absorption. The acidic mucus likely maintained surface epithelium pH and protease activity. Within a phylogenetic context, the absence of a pyloric caeca in G. pictus while possessing an intestine implies that this species is affiliated to groups that had branched off earlier than basal teleosts. Inc.

Microanatomy and ultrastructure of the foot of the infaunal bivalve Tegillarca granosa (Bivalvia: Arcidae)

In this study, the morphology and ultrastructure of the foot of Tegillarca granosa was compared with the bivalves from different habitats. The sediment of habitat of T. granosa is mostly a mixture of sand (68.93%) and mud (24.12%). The foot is wedge-shaped with multiple projections on the surface and covered with ciliary tufts. The epithelial layer is simple and composed of ciliated columnar epithelia and mucous cells. Although the mucous cells are distributed mostly in the epithelial layer, they are developed even in the connective tissues and muscle layers, and the mucous cells mostly contain acidic carboxylated mucosubstances. From the TEM observation, secretory cells are classified into three types. Type A secretory cell has a goblet form and is most widely distributed among the three types. Type B secretory cell has an oval form and the secretory granule has fibrous substance. Type C secretory cell has an elongated elliptic form and membrane-bounded secretory granules. The muscle fiber bundles are composed mainly of smooth muscle fibers. The smooth muscle fibers can be divided into two types. Type A muscle fibers have evenly distributed thick microfilaments between the thin microfilaments of cytoplasm. Type B muscle fiber has cluster of condensed microfilaments in the medulla cytoplasm while the cortical cytoplasm has loose distribution of thin microfilaments.

Ultrastructure and Glycoconjugate Pattern of the Foot Epithelium of the AbaloneHaliotis tuberculata(Linnaeus, 1758) (Gastropoda, Haliotidae)

The foot epithelium of the gastropod Haliotis tuberculata is studied by light and electron microscopy in order to contribute to the understanding of the anatomy and functional morphology of the mollusks integument. Study of the external surface by scanning electron microscopy reveals that the side foot epithelium is characterized by a microvillus border with a very scant presence of small ciliary tufts, but the sole foot epithelium bears a dense field of long cilia. Ultrastructural examination by transmission electron microscopy of the side epithelial cells shows deeply pigmented cells with high electron-dense granular content which are not observed in the epithelial sole cells. Along the pedal epithelium, seven types of secretory cells are present; furthermore, two types of subepithelial glands are located just in the sole foot. The presence and composition of glycoconjugates in the secretory cells and subepithelial glands are analyzed by conventional and lectin histochemistry. Subepithelial glands contain mainly N-glycoproteins rich in fucose and mannose whereas secretory cells present mostly acidic sulphated glycoconjugates such as glycosaminoglycans and mucins, which are rich in galactose, N-acetyl-galactosamine, and N-acetyl-glucosamine. No sialic acid is present in the foot epithelium.

Development of the larval anterior neurogenic domains of Terebratalia transversa (Brachiopoda) provides insights into the diversification of larval apical organs and the spiralian nervous system.

BackgroundLarval features such as the apical organ, apical ciliary tuft, and ciliated bands often complicate the evaluation of hypotheses regarding the origin of the adult bilaterian nervous system. Understanding how neurogenic domains form within the bilaterian head and larval apical organ requires expression data from animals that exhibit aspects of both centralized and diffuse nervous systems at different life history stages. Here, we describe the expression of eight neural-related genes during the larval development of the brachiopod, Terebratalia transversa.ResultsRadially symmetric gastrulae broadly express Tt-Six3/6 and Tt-hbn in the animal cap ectoderm. Tt-NK2.1 and Tt-otp are restricted to a central subset of these cells, and Tt-fez and Tt-FoxQ2 expression domains are already asymmetric at this stage. As gastrulation proceeds, the spatial expression of these genes is split between two anterior ectodermal domains, a more dorsal region comprised of Tt-Six3/6, Tt-fez, Tt-FoxQ2, and Tt-otp expression domains, and an anterior ventral domain demarcated by Tt-hbn and Tt-NK2.1 expression. More posteriorly, the latter domains are bordered by Tt-FoxG expression in the region of the transverse ciliated band. Tt-synaptotagmin 1 is expressed throughout the anterior neural ectoderm. All genes are expressed late into larval development. The basiepithelial larval nervous system includes three neurogenic domains comprised of the more dorsal apical organ and a ventral cell cluster in the apical lobe as well as a mid-ventral band of neurons in the mantle lobe. Tt-otp is the only gene expressed in numerous flask-shaped cells of the apical organ and in a subset of neurons in the mantle lobe.ConclusionsOur expression data for Tt-Six3/6, Tt-FoxQ2, and Tt-otp confirm some aspects of bilaterian-wide conservation of spatial partitioning within anterior neurogenic domains and also suggest a common origin for central otp-positive cell types within the larval apical organs of spiralians. However, the field of sensory neurons within the larval apical organ of Terebratalia is broader and composed of more cells relative to those of other spiralian larvae. These cellular differences are mirrored in the broader spatial and temporal expression patterns of Tt-FoxQ2 and Tt-otp. Corresponding differences in the expression of Tt-hbn, Tt-NK2.1, and Tt-FoxG are also observed relative to their respective domains within the cerebral ganglia of spiralians. Based on these data we argue that the anterior region of the bilaterian stem species included Six3/6, NK2.1, otp, hbn, fez, and FoxQ2 expression domains that were subsequently modified within larval and adult neural tissues of protostome and deuterostome animals.