Ultrastructural Aspects of Spicule Formation in the Solitary Ascidian Herdmania momus (Urochordata,Ascidiacea)

The solitary stolidobranch ascidian Herdmania momus contains numerous calcium carbonate spicules in its tunic and body tissues. The slender body spicules form inside complex sheaths in the body wall and branchial basket, where they remain for the life of the animal. The much smaller tunic spicules form inside the tunic blood vessels and then migrate to the tunic surface, where they become anchored by their spiny base. This paper is an ultrastructural investigation of the formation of the body spicules; the tunic spicules, which apparently form quite differently, will be the focus of a future study. The body spicules are composed of rows of closely packed acicular spines which form completely extracellularly. The spine tips are covered by flattened, highly pseudopodial sclerocytes bound together by tightly interdigitating cell processes. The basal regions of contiguous spines are covered by very thin sclerocyte cell processes. An organic matrix is present within the spines; its exact nature is not clear. A very dense extracellular inter-spine matrix is located between the spine tips and the contiguous basal regions. Presclerocytes within the sheaths between the spicules are probably responsible for formation of the extracellular structures of the sheaths. The presclerocytes appear to aggregate and transform into sclerocytes at the apical end of the spicule. New spines are added at the apical end of the spicule as well as between larger spines. Comparisons are made between body spicule formation in H. momus and skeletogenesis in echinoids.     

Pigmentation and tunic cells in Cystodytes dellechiajei (Urochordata, Ascidiacea)

The tunic of Cystodytes dellechiajei (Poly- citoridae), a colony-forming species of the Ascidiacea that contains biologically active alkaloids, was investigated using light microscopy, laser-scanning microscopy and nuclear magnetic resonance techniques. The colonies contain numerous individual zooids, which are embedded in a common tunic. Each zooid is protected by a firm capsule of overlapping calcareous spicules. The colonies lack blood vessels in the tunic, but six morphologically different types of tunic cells were found: pigment cells, bladder cells, vacuolated filopodial cells, granular filopodial cells, morula cells and granular cells. Rod-like bacteria were found in the tunic matrix. Bladder cells and pigment cells could be identified as storage units for acid and pyridoacridine alkaloids, making the tunic inedible and repelling predators. Filopodial cells have long filopodia, which probably are connected to each other. They may be involved in transportation processes within the tunic tissue. The functions of the morula cells and the granular cells are unknown as yet. With its several specialised cells, the tunic of C. dellechiajei represents a dynamic living tissue containing biologically active compounds. Accepted: 20 September 2000     

Tunic cells in pyrosomes (Thaliacea, Urochordata): cell morphology, distribution, and motility

Abstract. Cellular components of the tunic were histologically examined in 3 pyrosome species representing all 3 genera of the order: Pyrosoma atlanticum, Pyrosomella verticillata , and Pyrostremma spinosum . Three cell types are distributed in the tunic. Tunic amebocytes, irregularly shaped and motile, often contain granules and/or phagosomes. Spherical tunic cells contain many round vesicles with eosinophilic and acidic materials. Tunic net cells form a cellular network in which their long filopodia connect with one another. The net cells are densely distributed just beneath the tunic surface lining the common cloacal cavity and may produce tension to maintain the colony shape. The presence of net cells suggests a phylogenetic relationship between pyrosomes and some aplousobranch ascidians. Test fibers are multicellular cords that run in the tunic and connect the zooids. In P. atlanticum , they are attached to they are attached to the epidermal cells of the zooids, and transverse cloacal muscles are attached to the other (proximal) side of the epidermal cells. The test fibers may mediate coordination of the zooids and control muscle contraction.     

The morphology of allograft rejection in Styela plicata (Urochordata: Ascidiacea)

Summary This study identifies three discrete processes responsible for the rejection of tunic tissue transplanted between individuals of the solitary ascidian Styela plicata. The first stage of rejection is characterized by the destruction of blood vascular components within incompatible allografts. In the second phase, dense boundaries of extracellular material are deposited between grafts and the surrounding host tunic, effectively amputating the transplanted tissues. Finally, detached transplants undergo a gradual necrosis which results in the total degeneration of extracellular graft matrices. Of these three phases, the initial cellular depletion of allografts is responsible for the immunological specificity that is characteristic of histocompatibility in S. plicata. The subsequent amputation and necrosis of extracellular graft matrices are taken to be non-specific consequences of the initial cellular reactivity.     

Recent introduction of the dominant tunicate,Pyura praeputialis (Urochordata,Pyuridae) to Antofagasta,Chile

The large sessile tunicate Pyura stolonifera (Pleurogona: Stolibranchiata: Pyuridae), has been regarded as a complex taxon with disjointed distributions, including Australia (Pyura stolonifera praeputialis), South Africa (Pyura stolonifera stolonifera) and South America (Chile, Antofagasta: Pyura sp., the 'piure de Antofagasta'), and has been cited under at least five taxonomic combinations. The 'piure de Antofagasta' is a competitively dominant species in rocky intertidal habitats and shows a limited geographical range (60-70 km) exclusively inside the Bay of Antofagasta. Using cytochrome oxidase I (COI) mitochondrial sequence data from Pyura specimens of the three taxa we tested whether the Chilean taxon represents: (i) a Gondwana relict; (ii) a more recently divergent species; or (iii) a recently introduced species. The results suggest that the Chilean taxon is a recent introduction to Chile from Australian populations and that Pyura stolonifera praeputialis, from Australia, and the 'piure de Antofagasta' are geographical populations of a single species: Pyura praeputialis; whereas the South African taxon represents a second species: Pyura stolonifera.     

Tunic phagocytes are involved in allorejection reaction in the colonial tunicate Aplidium yamazii (Polyclinidae, Ascidiacea)

The colonial ascidian Aplidium yamazii exhibited an allorejection reaction when two allogeneic colonies were brought into contact at their growing edges or at artificial cut surfaces. This species has no vascular network in the tunic, unlike the botryllid ascidians, which have a vascular network throughout the colony's common tunic. In the allorejection reaction induced by contact at the growing edges, some small, hard-packed tunic masses were formed at the contact points. Histological and electron microscopic investigation of these tunic masses revealed that they contained aggregates of tunic cells, with tunic phagocytes being the major cell type present. Some of the tunic phagocytes in these tunic masses appeared to be disintegrating. When allogeneic colonies were placed in contact at their artificial cut surfaces, the colonies partially fused, then separated. In this allorejection reaction, some loosely packed tunic masses remained in the gap between the two withdrawn colonies. These results strongly suggest that the tunic phagocytes are likely to be the major effector cells in the allorejection reaction. We also propose that the tunic phagocytes are not only the effector cells in the allorejection reaction but also bear the sites of allorecognition.     

Occurrence of different secretin-like cells in the digestive tract of the ascidian Styela plicata (Urochordata,Ascidiacea)

Summary Secretin-like cells have been detected in the digestive tract of the ascidian Styela plicata by means of immunofluorescent and immunocytochemical methods.Especially, in the esophageal epithelium there are immunoreactive cells (S₂) in which a biogenic amine (5-HT) and a regulatory peptide (secretin) occur together. In the gastric epithelium only secretin-like cells (S₁) are present.Tests of cross-reactivity performed with glucagon, GIF and VIP, have confirmed the presence of a secretin-like molecule only in the S₁ and S₂ cells.     

Characterization and supramolecular architecture of the cellulose-protein fibrils in the tunic of the sea peach (Halocynthia papillosa,Ascidiacea, Urochordata)

Summary— The cellulose-protein fibrils, which constitute by far the bulk of the fibrous fraction of the sea peach tunic (Halocynthia papillosa), were structurally and chemically characterized, either in situ or after extraction procedures, with the use of classical electron microscoy combined with diffraction contrast imaging and electron diffraction, histochemistry, affinity cytochemistry and chemical analysis. These fibrils exhibit a cross-sectional shape close to a parallelogram. The cyrstallites forming their core, with lateral dimensions ranging from roughly 5 to 20 nm, are composed of native cellulose of higher crystallinity than that of plant cellulose. They are associated with acid mucopolysaccharidés (amps) and proteins which form a coating material appearing as a continuous sheath enveloping the axial crystallite in the cuticular layer or as patches more-or-less periodically distributed around and along the fibre axis in the fundamental layer. Tunicin, the alkali-insoluble fibrous fraction, is not pure cellulose, yielding only 22–60% of its dry weight as glucose equivalents, depending on the tunical layer. It is suggested that in addition to the high degree of crystallinity of the tunical cellulose, the presence of a significant amount of coating material composed of amino acids and proteoglycans firmly linked to cellulose molecules contributes to tunicin's high resistance to hydrolysis.     

An atrial membrane in the colonial ascidian, Ritterella tokioka Kott, 1992 (Urochordata: Ascidiacea) from Sagami Bay

An atrial membrane, similar to that previously reported (as a placental membrane) in Placentela crystallina Redikorzev, 1913, has been found in the holotype and other specimens of another aplousobranch ascidian, Ritterella tokioka Kott, 1992 (=R. pedunculata Tokioka, 1953). In serial sections, the membrane is seen to be an extensive fold of the atrial epithelium over the rectum that projects into the atrial cavity and supports developing embryos over its outer surface. The similar states in the atrial membrane seen in the two species are attributable to homoplasy, rather than homology.     

Tunic morphogenesis in the sea peach (Halocynthia papillosa,Ascidiacea, rochordata): Cellular events,the initiation of twisted fibrous arrangements and spine formation

Summary— The adult tunic of the sea peach (Halocynthia papillosa) shows a high degree of organisation. Tunic morphogenesis was monitored from the onset of tunic secretion until juveniles reached the age of 3 months. While some characteristics of the adult tunic are still missing, like certain types of intratunical cells and striated bodies, its main features have already developed by this time. Crucial events take place at or soon after the onset of metamorphosis (stage M 0). Cuticular spines cover the external surface of the juvenile. At least two types of intratunical cells enter the tunic and the fibrous material adopts a three-dimensional twisted helicoidal architecture. The initiation of this helicoidal arrangement of fibrils directly after stage M 0 is discussed regarding accompanying developmental events. The existence of cells that penetrate the outer compartment of the tunic at the end of larval life is reported for the first time.     

Possible Ascidian Counterpart to the Vertebrate Saccus Vasculosus with Reference to Pyura tessellata (Forbes) and Boltenia echinata (L.)

The morphology of the hitherto unknown larva of Pyura tessellata (Forbes) is described and the discovery of an auxiliary brain vesicle is presented. The vesicle was also found in another pyurid species Boltenia echinata (L.). The auxiliary vesicle communicates with the sensory vesicle at the level of the statocyte and is lined with cubodial epithelial cells carrying a 2 μm globular structure projecting into the lumen. These cells very closely resemble primitive tunicate coronet cells previously reported and coronet cells typical for the saccus vasculosus found in elasmobranchs and many ganoids and teleosts.     

Renewal of the gonads in Styela clava (Urochordata: Ascidiacea) as revealed by autoradiography with tritiated thymidine

DNA-synthesizing cells in the gonads of the ascidian Styela clava were labeled with tritiated thymidine and detected with autoradiography. In the testis, spermatogonia and primary spermatocytes are labeled after 1 hr. Labeled spermatozoa occur in the lumen of the testis follicles after 10 days and in the sperm ducts after 20 days. In the ovary, only germ cells (oogonia and pre-leptotene primary oocytes) and follicle cells are labeled after 1 hr. By 60 days, oocytes with basophilic cytoplasm (15–65 μ in diameter) are labeled; test cells embedded in larger eosinophilic oocytes (150 μ in diameter) are also labeled. Germ cells give rise to both oocytes and follicle cells. Through continued cell division, follicle cells give rise to test cells.     

Formation and ultrastructural organization of gonads in Oikopleura gracilis Lohmann, 1896 (Urochordata: Appendicularia)

This study deals with the formation and ultrastructural organization of the gonads in a common species of appendicularian, Oikopleura gracilis, from Peter the Great Bay. Light microscopy observations show that the gonads develop from a transparent primordium that is located in the basolateral part of the gonad cavity; the primordium increases in size in the process of development and differentiates into the testis and ovary. The testis is covered by a single layer of ultrastructurally uniform follicular epithelium and contains a population of proliferating male gonocytes. The ovary contains two types of germ line nuclei, which are large polyploid nuclei that belong to the auxiliary cells and small meiotic nuclei of the oocytes. The two nuclei types, together with a common cytoplasm, form a syncytium of the ovary, or the coenocyst. As in the dioecious Oikopleura dioica, the coenocyst of O. gracilis produces naked oocytes that are devoid of a type III follicular membrane. The coenocyst is covered by a single-layered follicular epithelium, in which two cell types can be distinguished ultrastructurally. Thus, the synchronous maturation of sex products in O. gracilis is achieved by the formation of the germ-line syncytium in the testis and the coenocyst in the ovary, which generates a large number of simultaneously ripening oocytes that are competent for fertilization.     

Styela clava Herdman (Urochordata, Ascidiacea), a successful immigrant to North West Europe: ecology, propagation and chronology of spread

Since its first occurrence at Plymouth, southern England, in 1952 the East Asiatic ascidianStyela clava has spread to many localities along the coasts of the south and west British Isles, Ireland, northern France, Belgium, the Netherlands, Denmark and Germany. While some dispersal may occur by natural means, spreading over long distances is probably due to transfer along with oysters when relaid elsewhere. Transport while attached to the hulls of ships or driftingSargassum is also possible.Styela clava is a large, hardy and fast-growing species with a tough, leathery tunic, and has no recorded enemies or native analogues among the NW European ascidian fauna. At many sites it has established dense populations of 500–1000 specimens/m² and in some cases has nearly outcompeted some of the native ascidian species.     

Origin and Differentiation of the Inner Follicular Cells during Oogenesis in Molgula pacifica (Urochordata), an Ascidian without Test Cells

In Molgula pacifica small previtellogenic oocytes are found between cells of the ovarian epithelium. Each oocyte subsequently grows within a compartment of the epithelium known as a primary follicle. The wall of the primary follicle is composed of outer follicular epithelial cells. While growing from about 15–70 μm in diameter, each oocyte gradually recruits a set of about 950 non-epithelial inner follicular cells. These cells co-differentiate in sets with each oocyte, but test cells never appear. The first filamentous components of the vitelline coat appear on the surface of an oocyte in places where it is in contact with undifferentiated (stage 2) inner follicular cells. Each fully differentiated inner follicular cell stores adhesive precursors in a large compartment of the endoplasmic reticulum and probably secretes components of the vitelline coat. There is no evidence that the outer follicular epithelial cells transform into inner follicular cells by dedifferentiation as has often been assumed. Inner follicular cells, in stage 1, are nearly identical to hemoblasts. Hemoblasts may form the inner follicular cells, but to do this they would have to cross the outer follicular epithelium and this phenomenon has not yet been seen.     

Fine Structure and Differentiation of the Acrosome-Like Structure in the Solitary Ascidians,Pyura haustor aud Styela plicata1

An acrosome-like structure has been recognized at the apex of mature spermatozoa of both Pyura haustor and Styela plicata. The acrosome-like structure of P. haustor is a slightly depressed ellipsoid, approximately 90 nm × 80 nm × 50 nm, in length, width and height, respectively, while that of S. plicata is an antero-posteriorly elongated, flattened vesicle, approximately 200 nm × 100 nm × 50 nm, in length, width and height, respectively. During spermiogenesis, two vesicles (50–80 nm in diameter) are found in a blister at the apex of early spermatids of both species. These vesicles, presumably derived from the Golgi apparatus, contain moderately electron-dense material. In late spermatids, these two vesicles appear to fuse to form an acrosome-like structure. Because of its extremely reduced size and the paucity of its contents, it is unlikely that the acrosome-like structure of these sperm contain a significant amount of chorion lysin(s). A well developed Golgi apparatus and many Golgi vesicles of various sizes are found in the cytoplasm of spermatids in both P. haustor and S. plicata. It is hypothesized that ascidian spermatozoa contain a poorly developed acrosome, and that the chorion lysin(s) are intercalated into the plasmalemma enclosing the sperm head.     

Variability in the diet of New Zealand sea lion (Phocarctos hookeri) at the Auckland Islands, New Zealand

We examined the stomach contents of 121 New Zealand (NZ) sea lions ( Phocarctos hookeri ) caught by the squid fishery during the summer/autumn 1997–2006 around the Auckland Islands (51°S, 166°E). Dietary variation was assessed among juveniles, lactating females, nonlactating females and males, and between areas on the Auckland Islands shelf. The digested fraction of the contents consisted mostly of opalfish ( Hemerocoetes spp.) (50.1% by number [ N ], 4.7% by mass [ M ]), rattail ( Coelorinchus spp.) (12.0% N , 2.4% M ), arrow squid ( Nototodarus sloani ) (14.1% N , 17.9% M ), octopus ( Enteroctopus zealandicus ) (2.1% N , 27.8% M ), and red cod ( Pseudophycis bachus ) (3.8% N , 4.3% M ). Opalfish was found in greater proportions in the stomachs of females (lactating: 58.1% N , nonlactating: 62.4% N ) and juveniles (56.9% N ) than males (14.5% N ). Juveniles caught smaller opalfish and rattail than adults did. Over all classes, sea lions ate larger prey in the east than in the north of the Auckland Islands shelf. The common prey—arrow squid and rattail—constitute an abundant resource at the edges of the Auckland Islands shelf, where lactating NZ sea lions forage. Although these key areas are far from the rookeries and impacted by the squid fishery, they may provide the only reliable resource able to support the cost of benthic foraging behavior in the deepest diver of all otariids.     

Sensory endings of the ascidian static organ (Chordata,Ascidiacea)

Summary The ultrastructure of the static organ is examined in larvae of Diplosoma macdonaldi, a colonial ascidian, and Styela plicata, a solitary ascidian; the results are similar. As previous workers found, the cell body of a unicellular statocyte lies in the lumen of the sensory vesicle and contains the statolith. A narrow neck connects the cell body to an anchoring foot in the floor of the sensory vesicle. Two previously undescribed sensory endings project into the lumen just to the left of the statocyte, one anterior and one posterior to the neck. A network of fine processes from each ending contacts the statocyte body. It is proposed that movements of the statocyte cell body are detected by these endings. They arise from neurons in the ventral wall of the sensory vesicle that project axons to the visceral ganglion. The placement of the sensory endings may allow discrimination of the directon of statocyte deflection.Abbreviations ax axons - bb ciliary basal body - bl basal lamina - c cilium - cr striated ciliary rootlet - ec ependymal cells - en endoderm - h hemocoel - ly lysosome - mv microvilli - n neuron - nf neurofilaments - ns neck of the statocyte - sb statocyte cell body - sd sensory dendrite - sn sensory neuron - sp sensory processes - stf statocyte foot - svl sensory vesicle lumen - zo zonula occludens