Endocrine cells in the gut of the ascidian Styela clava

Summary Ultrastructural studies have shown the presence of two types of granulated endocrine cell in the gut of Styela clava. Type I, which occurs in the stomach and intestine contains small irregular granules, each with a distinct halo. Type II, found only in the oesophagus contains larger rounded granules, often with little or no halo. The characteristics of these two cell types are compared with those of endocrine cells found in the digestive tracts of other protochordates and discussed with special reference to the evolution of gastrointestinal endocrine cells in vertebrates.The authors are grateful to Mr. R. Jones for photographic assistance. Animals were collected by courtesy of the Admiralty Marine Trials Station, Portsmouth, This research was carried out during the tenure of S.R.C. grant no. B/RG 82919 to one of us (M.C.T.). The localization of polypeptide hormones in the pharynx and gut of protochordates     

Calcitonin-like cells in the pharynx of the ascidian Styela clava

Summary A small group of granulated endocrine cells have been described in the endostylar region of the pharynx in Styela. These cells are argyrophilic and exhibit calcitonin-like immunofluorescence. Tests with antisera to other peptides all proved negative. Cells from animals exposed to elevated calcium levels showed a degree of degranulation. The possibility that these cells may represent ancestral ultimobranchial C cells is discussed.The authors are very grateful to Julia Polak and Susan Van Noorden, Royal Postgraduate Medical School, London, for the kind gift of all antisera used. Animals were collected by courtesy of Mr. David Houghton and his staff of the Admiralty Marine Trials Station, Portsmouth. This research was carried out during the tenure of SRC grant no. B/RG 82919 to one of us (M.C.T.) — The localization of polypeptide hormones in the pharynx and gut of protochordates     

Physiological energetics of the ascidian Styela clava in relation to body size and temperature

The ascidian Styela clava is widely distributed in northern China and is thought to be important in the functioning of estuarine systems. This ascidian may experience highly variable physiological conditions, and its physiological responses to these are of interest considering its ecological role. This study presents data on the physiological parameters in relation to body size and temperature (12-28 degrees C) of S. clava. Respiration, excretion, feces, ingestion and absorption energy were positive related to size and its mass exponents (b-values) varied from 0.2930 to 0.7488. Respiration energy increased with increasing, but critical values were found at 20 degrees C for energy of ingestion and absorption, while 24 degrees C for energy in feces and excretion. At the range of 16-24 degrees C, the scope for growth, gross and net growth efficiencies of ascidians increased with increasing temperature. The energy budget equations of different sized ascidians were obtained at different temperatures. Excretion energy shared a minimal fraction in ingestion energy (1.30-2.47%), the ratios of feces energy and respiration energy to ingestion energy were 46.53-64.27% and 10.26-80.75%, respectively. The physiological data obtained in the present study indicated that S. clava could adjust its energy budget according to the environment and its physiological conditions to meet their nutritional and energetic demands. In the range of experimental temperature (12-28 degrees C), 16-20 degrees C was suitable for the rearing of S. clava to achieve optimum development, while deficient metabolic adjustment induced a negative scope for growth of S. clava at 28 degrees C.     

Physiological energetics of the ascidian Styela clava in relation to body size and temperature.

The ascidian Styela clava is widely distributed in northern China and is thought to be important in the functioning of estuarine systems. This ascidian may experience highly variable physiological conditions, and its physiological responses to these are of interest considering its ecological role. This study presents data on the physiological parameters in relation to body size and temperature (12-28 degrees C) of S. clava. Respiration, excretion, feces, ingestion and absorption energy were positive related to size and its mass exponents (b-values) varied from 0.2930 to 0.7488. Respiration energy increased with increasing, but critical values were found at 20 degrees C for energy of ingestion and absorption, while 24 degrees C for energy in feces and excretion. At the range of 16-24 degrees C, the scope for growth, gross and net growth efficiencies of ascidians increased with increasing temperature. The energy budget equations of different sized ascidians were obtained at different temperatures. Excretion energy shared a minimal fraction in ingestion energy (1.30-2.47%), the ratios of feces energy and respiration energy to ingestion energy were 46.53-64.27% and 10.26-80.75%, respectively. The physiological data obtained in the present study indicated that S. clava could adjust its energy budget according to the environment and its physiological conditions to meet their nutritional and energetic demands. In the range of experimental temperature (12-28 degrees C), 16-20 degrees C was suitable for the rearing of S. clava to achieve optimum development, while deficient metabolic adjustment induced a negative scope for growth of S. clava at 28 degrees C.     

A cytochemical and immunofluorescence study of endocrine cells in the gut of the ascidian Styela clava

Summary Strong secretin-like immunofluorescence has been demonstrated in endocrine-like cells from the gastric epithelium of Styela. These cells also stain with lead haematoxylin and exhibit a brilliant formaldehyde-induced fluorescence, but do not show any other cytochemical features characteristic of the mammalian APUD series. Tests with antisera to glucagon, gastrin and somatostatin all proved negative. In the oesophagus tests with all four antisera proved negative. The significance of these results is discussed in relation to the phylogeny of vertebrate gastro-intestinal hormones.     

Effect of the ascidian Styela clava on the growth of small holothurians of the species Apostichopus japonicus

We studied the effect of the common ascidian Styela clava on the growth of small holothurians of the species Apostichopus japonicus (Selenka, 1867). In a traditional aquaculture system, the oxygen consumption rates, ammonia excretion rates, and ingestion rates of small A. japonicus were reduced significantly, which suggested that this group was stressed by the presence of large A. japonicus, and that this stress grew stronger with time. Oxygen consumption rates, ammonia excretion rates, and ingestion rates of small A. japonicus in an integrated aquaculture (IA) system with S. clava, microalgae, and A. japonicus were higher than those observed in the traditional culture system. Metabolic and digestive enzymes were more active in small A. japonicus in the IA system than in those in the traditional aquaculture system. These results suggest that the IA model did not affect the growth of large A. japonicas, but produced a marked positive impact on the growth of small individuals.     

Tolerance of the invasive tunicate Styela clava to air exposure

Styela clava is a subtidal invasive marine species in Northern Europe, Atlantic Canada, Australia and New Zealand. It grows attached to solid substrata, including boat hulls, ropes, moorings, piers and aquaculture equipment, all of which can aid its spread to new locations. It interferes with feeding of mussels and oysters, and increases their harvesting costs. Being subtidal, it could be assumed that tunicates would rapidly die in air and thus exposure to air would be a practical method to prevent their spread on boats and equipment. This study tested their survival when exposed to air for up to (1) 120?h at a constant temperature of 10?°C, (2) shade ambient 15–27?°C, and (3) full sun ambient 15–29?°C. Humidity was consistently high (78–100%). The results indicated that survival was longer when the air temperature was cooler. Larger individuals of S. clava generally survived for longer out of seawater than smaller individuals. The results predict that two weeks of exposure to air for two weeks could be an effective management method to eradicate S. clava from marine equipment when the air temperature is 10?°C. However, drying time would be less under conditions of low humidity and under direct sunlight.     

Purification and characterization of a humoral opsonin from the solitary urochordate Styela clava.

1. We have previously identified opsonic activity in the plasma of the solitary urochordate, Styela clava. 2. Here, we report the purification and further characterization of the opsonic molecule. 3. Two purification methods were employed. 4. Gel filtration yielded one strongly opsonic fraction that contained a single, electrophoretically-resolved protein. 5. Opsonic activity was dose-dependent and sensitive to tryptic digestion and heat denaturation. 6. SDS-PAGE and calibrated gel filtration indicated the opsonic protein was a 17.5 kDa monomer while isoelectrofocusing indicated a single pI of 7.0. 7. In an alternative procedure, a similar opsonic activity and protein were isolated by affinity purification using whole yeast cells.     

Endocrine Cells in the Oesophagus of the Ascidian Styela clava,a cytochemical and immunofluorescence study

Summary Immunocytochemical studies have demonstrated the occurrence of an insulin-immunoreactive cell type in the oesophageal epithelium of the Ascidian Styela clava. Staining with aldehyde fuchsin has demonstrated a number of similar small, triangular, cells located on the basement membrane, which may have an endocrine function. Argyrophilic cells have also been found, suggesting the presence of a second endocrine cell type. The absence of argentaffin cells has led us to believe that the cells so far observed do not produce biogenic amines such as 5-HT (5-Hydroxytryptamine). The nature of these cells is discussed with reference to endocrine-like cells found in the digestive tracts of other protochordates.Animals were collected by courtesy of the Admiralty Marine Trials Station, Portsmouth. This research was carried out during the tenure of an S.R.C. grant no. B/RG 82919 to one of us (M.C.T.). —The localisation of polypeptide hormones in the pharynx and gut of protochordates     

In vitro allogeneic cytotoxicity in the solitary urochordate Styela clava.

Hemocytes from the solitary urochordate Styela clava can effect allogeneic cytotoxicity in vitro. Spectrophotometric and microscopic quantification of eosin-y dye exclusion revealed significantly greater frequencies of cell death in allogeneic hemocyte cultures when compared to autogeneic controls. This cytotoxic response was characterized by 1) transient activity such that specific cytotoxicity could be detected for only 4 hours of culture though continued specific killing may have been obscured by spontaneous cell death; 2) a necessity for cellular interaction demonstrated by the elimination of allogeneic cytotoxicity in the absence of cell contact; 3) killing of multiple targets by effector cells due to high levels of response at low allogeneic ratios; 4) insensitivity to altered temperature; 5) increased cytotoxicity in the absence of autologous plasma; 6) an absence of xenogeneic reactivity; 7) the presence of three hierarchical levels (low, intermediate, and high) of response. These data reflect events involved in the recognition of allogeneic cellular determinants resulting in specific cytotoxicity effected by immunocompetent cells. Such in vitro recognition and cytotoxic recognition and cytotoxic reactivity may be responsible for adaptive reactions caused by histoincompatibility in solitary tunicates.     

The sequence of a sea urchin muscle actin gene suggests a gene conversion with a cytoskeletal actin gene

Summary We report the nucleotide sequence of the single muscle actin gene of the sea urchinStrongylocentrotus purpuratus. Comparison of the protein-coding sequence of this muscle actin gene (pSpG28) with that of two linked sea urchin cytoskeletal actin genes (pSpG17 and CyIIa) reveals a region of exceptional sequence conservation from codon 61 through codon 120. Furthermore, when silent nucleotide changes are compared, the conservation of this region is still evident (7.9% silent site differences in the conserved region vs 43.3% silent site differences in the rest of the gene when pSpG28 and CyIIa are compared), indicating that the conservation is not due to particularly stringent selection on the portion of the protein encoded by this region of the genes. These observations suggest that a gene conversion has occurred between the muscle actin gene and a cytoskeletal actin gene recently in the evolution of the sea urchin genome. Gene conversion between nonallelic actin genes may thus play a role in maintaining the homogeneity of this highly conserved gene family.     

Protochordate immunity--II. Diverse hemolymph lectins in the solitary tunicate Styela clava

Hemolymph lectins (agglutinins) of the tunicate Styela clava were analyzed by agglutination, cross-absorption and carbohydrate-hemagglutination inhibition using several vertebrate erythrocytes. Lectin activity was heat labile, dependent on divalent cations and refractory to neuraminidase-treated erythrocytes. Four lectins with different carbohydrate specificities were found. Carbohydrate specificities included L-rhamnose, D-glucuronolactone, maltose, D-galactosamine, D-mannosamine, D-galactose, hyaluronic acid and bacterial lipopolysaccharide. Since S. clava lectins can be inhibited by carbohydrates found in the extracellular capsule or cell wall of most bacteria, we propose that the lectins may be part of the tunicate immuno-defense system.     

Fine- and regional-scale genetic structure of the exotic ascidian Styela clava (Tunicata) in southwest England, 50 years after its introduction

Styela clava , an ascidian native to the northwest Pacific, was first recorded in the Atlantic at Plymouth, southwest England, in 1953. It now ranges in the northeast Atlantic from Portugal to northern Denmark, and has colonized the east coast of North America. Within the region of first introduction, we aimed to characterize current genetic diversity in the species, elucidate the respective roles of human-aided vs. natural dispersal, and assess the extent of larval dispersal by looking for genetic differentiation at very small scales. Eight sites, mostly marinas, were studied along c . 200 km of coast in southwest England encompassing Plymouth. Five microsatellite loci were genotyped in 303 individuals to analyse gene flow at regional (among sites) and fine (within sites) scales. F -statistics and assignment tests were used to investigate regional genetic structure. At the fine scale, deviation from mutation–drift equilibrium was tested, and isolation by distance and genetic clustering analyses were undertaken. Significant genetic differentiation existed between sites, unrelated to geographical separation; migration between geographically distant marinas was inferred, highlighting the likely importance of human-mediated dispersal in range expansion and occupancy by S. clava . Fine-scale population structure was present within at least four sites, which may be explained by the limited dispersal ability of this ascidian and recruitment from differentiated pools of larvae. Populations in enclosed marinas had higher self-recruitment rates than those in open sites. Some marinas might therefore function as reservoirs of propagules for subsequent spread, whereas others might be sinks for migrants.     

Stress levels over time in the introduced ascidian Styela plicata: the effects of temperature and salinity variations on hsp70 gene expression

Species distribution, abundance, and long-term survival are determined by biotic and abiotic regimes. However, little is known about the importance of these factors in species range expansion. Styela plicata is a solitary ascidian introduced all over the world by ship fouling, including salt marsh habitats, where introduced populations must tolerate high seasonal variations in temperature and salinity. To determine the seasonal stress levels in a salt marsh population of S. plicata, we quantified heat shock protein (hsp70) gene expression using quantitative real-time PCR throughout a 2-year cycle. Results showed that hsp70 expression varied over time, with higher stress levels recorded in summer and winter. Periodic conditions of high temperatures, particularly when coupled with low salinities, increased hsp70 gene expression. Mortality events observed every year around June were concurrent with sharp increases in temperature (>6°C), indicating that drastic changes in abiotic factors may overwhelm the observed stress response mechanisms. Determining the ability of introduced species to cope with stress, and the thresholds above which these mechanisms fail, is fundamental to predict the potential expansion range of introduced species and design efficient containment plans.     

Temporal and spatial modulation of a cytoskeletal antigen during peripheral axonal pathfinding.

A neuron-specific cytoskeletal antigen (5E10), whose expression pattern during initial motoneuron outgrowth into the chick limb suggests that it is playing a role in axon guidance, is described. This antigen, which was shown to be a phosphorylated epitope, probably of the intermediate weight neurofilament protein (NF-M), exhibits a highly stereotyped and spatially heterogeneous pattern of expression. The point of onset of expression, which was abrupt and occurred in the distal axon and base of the growth cone, differed between groups of neurons that projected to different targets. Specifically, expression occurred from positions where previous perturbation experiments suggested that the axons in question would begin responding to specific guidance cues, and it remained high along the axon from this point to the target. Expression of this antigen could also be induced in cultured motoneurons by activating several second messenger systems.