A Rostral Sensory Mechanism in Oikopleura dioica (Appendicularia)

The ventral sense organ, below the mouth, is composed of 30 primary sensory cells situated in a row perpendicular to the long axis of the animal. Each cell carries one long and slender, modified cilium which arises from an apical pocket in the cell. The sensory cells project 15 axons at each side of the pharynx to the brain, which is rostrally paired and terminates in bulb-like swellings. Each of these bulbs contains four cell bodies, which, according to their fine structure, as well as the synaptic connections with receptor and brain fibers, belong to three different types. It is suggested that the sense organ is a chemosensor and that its remarkable similarity to the vertebrate mechanisms for olfaction makes it probable that the appendicularian Oikopleura dioica possesses a ‘protochordate’ counterpart to the craniate olfactory apparatus.     

The Oral Gland Cells of Oikopleura dioica (Tunicata Appendicularia)

Light and electron microscopic studies showed that the oral gland cells have two quite different zones. Medially, the basal zone is in contact with body fluids and the endostyle. Its strongly pyroninophile cytoplasm contains the extremely digitated nucleus and numerous small mitochondria. Laterally, the apical zone contacts the epidermis and it may also send a process between epidermal cells and deliver cell fragments into the primordium of the new house. This cell zone contains numerous membranes. It is concluded that the oral gland cells are light producing glands and that the membrane-rich cell fragments which are incorporated into the house wall are the source of the bioluminiscence which has been reported from empty houses. The ontogenetically related subchordal cells have a similar structure and it is possible that also these cells are light producers.     

Primitive Coronet Cells in the Brain of Oikopleura (Appendicularia,Tunicata)

Abstract A peculiar cell type is described from the sensory vesicle of the brain of the appendicularian tunicate Oikopleura dioica. The cells carry globular, modified cilia and resemble in several other respects the coronet cells of Saccus vasculosus in many fishes. This finding, together with earlier reports of similar cells in the sensory vesicle of ascidian tadpoles, makes it probable that a primitive form of coronet cells is a regular constituent of the tunicate brain cavity. The existence of coronet cells in the tunicate line of evolution is of interest from a phylogenetical as well as functional point of view. The author proposes that the tunicate sensory vesicle corresponds to part of the craniate third ventricle and that the tunicate coronet cells are involved in the regulation of the ventricle fluid composition.     

Distribution of peroxidase and iodination activity in the endostyles of Oikopleura albicans and Oikopleura longicauda (Appendicularia,Chordata)

Summary Oikopleura albicans and O. longicauda belong to the two subgenera Vexillaria and Coecaria, respectively. The morphology and ultrastructure of their endostyles were investigated with conventional microscopic procedures as well as with DAB cytochemistry and ¹²⁵I autoradiography at both light- and electron-microscopic levels. As expected, the general morphology of these endostyles is similar to all hitherto examined endostyles. They possess a ventral portion consisting of alternating glandular and ciliated cell zones, probably serving food capture, and a dorsal region, the corridor. Autoradiographic grains were found mainly in the corridor lumen associated with the apical surface of the two central rows of corridor cells. The same cells also gave strong positive reactions for peroxidase, the iodinating enzyme. Peroxidase activity was found in the apical plasma membrane as well as in the nuclear envelope, rough endoplasmic reticulum, Golgi area and cytoplasmic vesicles. Definitive conclusions concerning an apical uptake and subsequent release into the body fluid of iodinated material could not be made from the present experiments. Our investigations indicate that the two central rows of corridor cells in both subgenera of oikopleurids constitute the protothyroid region, possibly homologous to the vertebrate thyroid gland.     

House Production by Oikopleura dioica (Tunicata, Appendicularia) Under Laboratory Conditions

We examined the generation time and the house renewal rate ofOikopleura dioica under various conditions. Animals were fedtwo flagellates, Isochrysis galbana and Tetraselmis sp., withconcurrent determination of the carbon contents of body andhouse to estimate house production. The generation time was6 days at 15°C, 4 days at 20°C and 3 days at 25°Cat both 25 and 30 p.s.u. with a food concentration of 4 x 10⁴cells ml^–1. The carbon content of newly secreted housesranged from 0.5 to 0.8 µg, corresponding to 15.3 ±4.8% of body carbon. The house renewal rates increased withincreasing temperature and decreasing salinity. Food concentrationsranging from 100 to 16 x 10⁴ cells ml^–1, body size andlight condition had no effect on house renewal rate. Cloggingof the inlet filter by adding the large diatom Ditylum sol causedan increase in house renewal rates. The total number and carboncontent of houses during an animal's lifetime ranged from 46to 53 houses and from 6.5 to 10.4 µg, respectively. Sincedaily house production calculated for the O. dioica populationcorresponded to 130–290% of its biomass and daily discardedhouse materials corresponded to 490–1100% of the biomass,this organism must play an important role as a producer of macroscopicaggregates.     

The house of Oikopleura dioica (Tunicata,Appendicularia): Structure and functions

Summary Using appropriate techniques, we have studied in detail the structure and functions of the house of the appendicularian Oikopleura dioica Fol, 1872. In particular, we describe the structure of the inlet funnel, the reinforcing chamber on each side of the tail chamber and its function, the role and the structure of the valves located between the dorsal and water exit chambers and the structure and the role of the exit sphincter. In conclusion, we show that in addition to filtration of particles, movement of the tail also helps location of a favorable particulate environment in the sea.     

The Ciliated Brain Duct of Oikopleura dioica (Tunicata,Appendicularia)

Transmission electron microscopy revealed varying morphology of the dorsal part of the duct indicating the occurrence of different stages of activity. In some animals the tip cavity contains an amorphous substance which, at another phase of activity, is released into the surrounding hemocel through interstices in the walls of the tip. A transitional zone separates the dorsal tip from the large ventral part of the duct, where the wall, near the transitional zone, consists of a single layer of thin cells joined by tight junctions. Ventrally, the large ciliated cells lie in two levels near the opening into the buccal cavity. They bear numerous, tightly packed cilia which beat vigorously in living animals. The organization of this part resembles that of protonephridia. An excretory function is suggested for this part of the duct.     

Production of Oikopleura dioica (Appendicularia) following a picoplankton 'bloom' in a eutrophic coastal area

The ecological importance of the appendicularian Oikupleuradioica as a picoplankton predator and a metazoan secondary producerwas assessed in summer 1995 in the Seto Inland Sea, Japan. Justafter the collapse of a red tide of Gymnodinium mikimotoi, theabundances of bacteria and picocyanobacteria increased rapidlyand attained values of     

Productivity and grazing impact of Oikopleura dioica (Tunicata, Appendicularia) in Tokyo Bay

To estimate the productivity and grazing impact of a commonappendicularian species Oikopleura dioica in Tokyo Bay, monthlyobservations on its abundance and vertical distribution wereconducted during 2000. The abundance peaked in February andOctober, but was low during summer. Seasonal fluctuations inproductivity were similar to those of the abundance, with maximumvalues of 92, 134 and 508 mg C m^–2 day^–1 for somatic,new house and discarded house productivity in October and 206mg C m^–2 day^–1 for fecal pellet productivity inFebruary, respectively. The averaged biomass of O. dioica wasonly 2.5% of that of copepods; however, the secondary productivityof the former corresponded to 12.4% of the latter. Daily grazingimpact on particulate organic carbon ranged from 0.05% to 5%,which is close to the impact by copepods. These results implythat in Tokyo Bay, where small copepods and jellyfish are abundant,O. dioica is an important component of the ecosystem becauseit bridges between small primary producers and higher consumers.     

The Subchordal Cells of Oikopleura dioica and O. albicans (Appendicularia,Chordata)

Abstract Oikopleura dioica has two large subchordal cells which were studied in vivo and with light and electron microscopy. They have fixed positions within the haemocoel of the tail but change their morphology continually by protruding and withdrawing processes in an amoeboid manner. Also the fine structure varies considerably from one animal to the next. The cell surface sometimes indicates a strong pinocytotic activity with many coated pits and vesicles. In other cases there are many small vesicles which are interpreted as exocytotic. They are found both in the cytoplasm close to the plasma membrane and in the haemocoelic fluid. Subchordal cells with no pronounced surface activity have a large amount of rough endoplasmic reticulum, which suggests that they synthesize proteins. The presence of subchordal cells in the tail is correlated with the presence of the ontogenetically related bioluminescent oral gland cells in the pharynx; either both types exist simultaneously or both are lacking. It is speculated that the two cell types are also functionally co-ordinated. Oikopleura albicans has a multitude of tiny subchordal cells which have essentially the same fine structure as the two large Oikopleura dioica cells.     

A transmission electron microscopic investigation of the sensory vesicle in the brain of Oikopleura dioica (Appendicularia)

Summary The structure of three cell types in the sensory vesicle is described: (1) The statocyte, with its intracellular statolith, is attached to the medial wall of the vesicle via delicate shaft cells. (2) Cells along the dorsal, ventral and lateral walls which contact the surface of the statocyte with long, slender cilia. These cells are presumed to be primary sensory cells. (3) Presumed secretory cells, along the rostral and dorsal walls, may have a dual function: (a) secretion of the vesicle fluid, and (b) stabilization of the wall by turgor created in characteristic intercellular cavities. The sensory vesicle in Oikopleura contains undoubtedly typical statocyst components adequate for a free-swimming animal, whereas the ascidian system is suggested to be a device that responds to gravitational stimuli and, together with temporary photoreceptors, aids the larva in finding optimal settling conditions.     

The neural connection between the Langerhans receptor cells and the central nervous system in Oikopleura dioica (Appendicularia)

Summary Transmission electron microscopic analysis of serial sections showed that the receptor cells are innervated by only one neuron and not two as previously believed. The neuron's two dendrites constitute the afferent sensory nerves to the caudal ganglion where the neuron's cell body is located. Its neurite was traced a few micrometers, but the synaptic terminals were not identified. This sensory system in Oikopleura is compared with a similar caudal sensory system in the tadpole larva of Diplosoma macdonaldi investigated by Torrence and Cloney. Wiring diagrams are proposed for the two systems. The ganglia, which receive the afferent sensory neurons, are discussed in terms of models for further research on simple integration systems.     

Lipid composition of the pelagic tunicate Dolioletta gegenbauri (Tunicata, Thaliacea)

The free-swimming sexual stage (gonozoid) of the doliolid, Doliolettagegenbauri, was abundant at a site in the NE Atlantic (58W,20N) during June 1996. Lipid analysis indicated that theseorganisms were comprised mainly of polar lipid with only lowlevels of neutral storage lipid (triacylglycerols) and thisis consistent with a life history strategy of rapid somaticgrowth and gamete production. Fatty acid analysis of the gonozoidssuggested a diverse microplanktonic diet, comprising diatoms,flagellates and dinoflagellates. The fatty acid compositionof D.gegenbauri was dominated by 20:5(n–3) and 22:6(n–3),and is of high nutritional value to predators, particularlyfish. The results are discussed in relation to the organisms'life history strategy and their potential nutritional impacton higher trophic levels.     

Simple procedure for sperm cryopreservation in the larvacean tunicate Oikopleura dioica

The larvacean tunicate Oikopleura dioica is an attractive organism for studies of the development, evolution, and physiology of chordates, showing considerable promise for genetic approaches given its short life cycle of five days. To facilitate future genetic studies, the development of protocols for the maintenance of individual strains is essential. Here we report a simple and practical protocol for the cryopreservation of sperm using liquid nitrogen (-196°C) and dimethyl sulfoxide (DMSO) as a protective agent. The quality of the frozen-thawed sperm was evaluated in terms of fertilizing ability and subsequent development of the fertilized eggs. We examined several parameters to optimize the efficiency of cryopreservation, such as the concentration of DMSO, the method for acclimation of sperm to DMSO before freezing, and for placing sperm in liquid nitrogen, as well as the pH of the seawater used in resuspending the thawed sperm. We confirmed that viable sperm were recovered after preservation for more than one year. In addition, mature animals, and even a subsequent generation, were obtained from eggs fertilized by the cryopreserved sperm. The present procedure seems to be simple and sufficiently practical for maintenance of future established lines of O. dioica using frozen sperm.     

Production of Oikopleura longicauda (Tunicata: Appendicularia) in Toyama Bay, southern Japan Sea

Oikopleura longicauda occurred throughout the year in ToyamaBay, southern Japan Sea, and analysis of its size compositionand maturity revealed that reproduction was continuous overtheyear. Somatic growth production (P_g) varied with season from0.03 to 103 mg carbon (C) m^–2day^–1 (annual P_g 4.5g C m^–2), and house production (P_e) from 0.11 to 266 mgC m^–2 day^–1 (annualP_e 11.3 g C m^–2). The annualP_g/B ratio was 176. Compared with production data of some predominantzooplankton species in Toyama Bay, it is suggested that despitetheir smaller biomass, appendicularians are an important secondaryproducer.     

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.     

Zooplankton growth rates: the larvaceans Appendicularia, Fritillaria and Oikopleura in tropical waters

The growth rates of Appendicularia sicula, Fritillaria borealissargassi, Fritillaria haplostoma, Oikopleura dioica and Oikopleuralongicauda were determined from microcosms incubated in situat 23C in Jamaican waters. Experiments were conducted fromoligotrophic offshore waters, through mesotrophic Lime Cay andeutrophic Kingston Harbour in both natural and nutrient-enhancedphytoplankton communities. Length-weight relationships werecalculated for two of these species: O.longicauda log W=2.47log TL –6.10 and F.haplostoma log W=2.44 log TL –7.37,where weight (W) is in micrograms and trunk length (TL) is inmicrometres. Instantaneous growth rates averaged 1.7–2.5day^–1 for the five species and were observed as high as3.3 day^–1 These instantaneous rates are equivalent todaily specific growth rates averaging 4.6–11.4 and rangingup to 28. In larger genera, growth rates were related positivelyto picoplankton and nanoplankton concentration, and negativelyto the biomass of larvaceans, but in the smallest species growthwas unrelated to these factors. However, because the variabilityin these two factors within microcosms exceeded their naturalrange of variability, growth rates of larvaceans may normallybe unlimited by resources or population density effects. ¹Present address :Monterey Bay Aquarium Research Institute 7700Sandholdt Road, Moss Landing, CA 95039-0628, USA ²Present address :Bedford Institute of Oceanography PO Box 1006,Dartmouth, Nova Scotia B2Y 4A2, Canada     

Fertilization in Oikopleura dioica (Tunicata,Appendicularia): Acrosome reaction,cortical reaction and sperm-egg fusion

Summary Fine structural changes in the egg and sperm are described during gamete interaction in Oikopleura dioica, an appendicularian tunicate. The unfertilized egg has a vitelline layer 80 nm thick and a perivitelline space about 5 m wide. In the peripheral cytoplasm are a few cortical granules 0.6×0.7 m in diameter and areas rich in parallel cisternae of rough endoplasmic reticulum alternating with areas rich in long mitochondria. In the deeper cytoplasm the predominant organelles are multivesicular bodies. From 25 s to 60 s after insemination, the egg transiently elongates, although with no obvious cytoplasmic rearrangement, and the egg surface becomes bumpy. During this interval sperm enter the egg, and the cortical granules undergo exocytosis. After expulsion into the perivitelline space, the cortical granule contents do not appear to change their shape or blend with the vitelline layer, which neither elevates further nor loses its ability to bind sperm. On encountering the egg, the sperm undergoes an acrosome reaction involving exocytosis of the acrosome and production of an acrosomal tubule. The acrosomal contents bind the sperm to the vitelline layer, and the posterior portion of the acrosomal membrane and the anterior portion of the nuclear envelope evaginate together to form an acrosomal tubule, which fuses with the egg plasma membrane to form a fertilization cone. By 45 s after insemination, the sperm nucleus, centriole, mitochondrion and at least the anterior portion of the axoneme are within the fertilization cone. By 60 s sperm entry is complete. In having eggs with a cortical reaction and sperm with an acrosome reaction, O. dioica resembles echinoderms and enteropneusts and differs markedly from ascidian tunicates, which lack both these features. The relatively unmodified pattern of gamete interaction in O. dioica in comparison with the highly modified pattern in ascidians is difficult to reconcile with the neoteny theory that appendicularians have evolved via ascidian ancestors. The present results are more consistent with the idea that an appendicularian-like ancestor gave rise to ascidians.     

Junctional complexes of the branchia and gut of the tunicate,Pyrosoma atlanticum (Pyrosomatida,Thaliacea)

Summary The branchia of pyrosomes has been found to be like that of ascidians in that it shares with the latter, besides the presence of peribranchial chambers, stigmata bordered by clusters of seven rows of flattened cells, each bearing a single row of long cilia. The intercellular junctions between ciliated cells of the branchial basket and between the cells lining the gut, in pyrosomes, have been studied in thin sections and by freeze-fracture. In both tissues tight and gap junctions are present, but the former are much more extensive in the gut. The gap junctions in the branchial basket exhibit some atypical features. Moreover, although there are extensive zonulae adhaerentes between the ciliated cells of the branchial basket, there are none between the epithelial cells of the intestinal tract. This feature of the branchiae, together with the alignment of their cells into highly ordered rows of seven cells, are similar to those found in some groups of ascidians. The evolutionary relationships between pyrosomes and the aplousobranch ascidians are considered in the light of these results.