Ultrastructure and embryonic development of a syconoid calcareous sponge

Abstract. Recent molecular data suggest that the Porifera is paraphyletic (Calcarea+Silicea) and that the Calcarea is more closely related to the Metazoa than to other sponge groups, thereby implying that a sponge‐like animal gave rise to other metazoans. One ramification of these data is that calcareous sponges could provide clues as to what features are shared among this ancestral metazoan and higher animals. Recent studies describing detailed morphology in the Calcarea are lacking. We have used a combination of microscopy techniques to study the fine structure of Syconcoactum Urban 1905, a cosmopolitan calcareous sponge. The sponge has a distinct polarity, consisting of a single tube with an apically opening osculum. Finger‐like chambers, several hundred micrometers in length, form the sides of the tube. The inner and outer layers of the chamber wall are formed by epithelia characterized by apical–basal polarity and occluding junctions between cells. The outer layer—the pinacoderm—and atrial cavity are lined by plate‐like cells (pinacocytes), and the inner choanoderm is lined by a continuous sheet of choanocytes. Incurrent openings of the sponge are formed by porocytes, tubular cells that join the pinacoderm to the choanoderm. Between these two layers lies a collagenous mesohyl that houses sclerocytes, spicules, amoeboid cells, and a progression of embryonic stages. The morphology of choanocytes and porocytes is plastic. Ostia were closed in sponges that were vigorously shaken and in sponges left in still water for over 30 min. Choanocytes, and in particular collar microvilli, varied in size and shape, depending on their location in the choanocyte chamber. Although some of the odd shapes of choanocytes and their collars can be explained by the development of large embryos first beneath and later on top of the choanocytes, the presence of many fused collar microvilli on choanocytes may reflect peculiarities of the hydrodynamics in large syconoid choanocyte chambers. The unusual formation of a hollow blastula larva and its inversion through the choanocyte epithelium are suggestive of epithelial rather than mesenchymal cell movements. These details illustrate that calcareous sponges have characteristics that allow comparison with other metazoans—one of the reasons they have long been the focus of studies of evolution and development.     

Die Aufnahme partikulärer Nahrung beiReniera sp. (Porifera)

The choanocyte chambers of the marine spongeReniera sp. protrude with their curved outer surface free into the incurrent canals. The water is sucked into the chambers by cavities between the choanocytes. Particles up to 1 µm in diameter may enter the chambers with the water current. These particles are trapped on the outer surface of the choanocyte collars and are ingested by the choanocytes and processes of the pinacocyte epithelium of the incurrent canal system, which project into the chambers. Bigger particles are retained in the incurrent canals mainly on the outer surface of the choanocyte chambers. They are ingested by pinacocytes of the canal wall and transported to cells of the mesenchyme. The present investigation shows the great importance of the pinacocyte epithelium of the incurrent canal system for suspension feeding inReniera sp.     

The filtration apparatus for food collection in freshwater sponges (Porifera,Spongillidae)

Summary The freshwater sponges (Spongillidae) feed by filtering out small particles from the water passing through them by means of strainer devices in the flagellated chambers. These are filamentous, fine-meshed structures at the distal ends of the choanocyte collars formed of a mucous material similar to that in the glycocalyx. Each strainer separates its flagellated chamber into an outer and an inner zone. The strainers are an extremely efficient filtering mechanism.     

Formation and construction of asexual buds of the freshwater spongeRadiospongilla cerebellata (Porifera,Spongillidae)

Summary The buds ofRadiospongilla cerebellata are formed asexually. Budding can be induced experimentally by injuring the sponge. The first sign of budding is a slight elevation of some surface areas, which proceed to rise rapidly so that they soon protrude conspicuously from the surface of the sponge. As a bud develops, the broad base joining it to the mother sponge narrows to a stalk, which finally breaks. The free buds drift in the water for 15–20 min and then settle, forming new sessile sponges. The buds, 1.5–2.5 mm in diameter, have an internal organization identical with that of the mother sponge. They are enclosed in a layer of pinacoderm perforated by dermal pores. Under the pinacorderm there is a shallow subdermal space, which is in communication with the incurrent canals leading to the choanocyte chambers. The water sucked into these chambers proceeds into the excurrent canal system and emerges from the sponge through the oscular tube. Spicules projecting radially from the bud bear apical tufts of microscleres. The skeletal spicules of the buds, like their choanocyte chambers, are smaller than those in the mother sponge. The chambers expand to their mature size by choanocyte mitosis. Buds and sponges are colored green by intracellular symbiotic algae of the genusChlorella.     

Hydrodynamics in early animal evolution

Choanoflagellates and sponges feed by filtering microscopic particles from water currents created by the flagella of microvillar collar complexes situated on the cell bodies of the solitary or colonial choanoflagellates and on the choanocytes in sponges. The filtering mechanism has been known for more than a century, but only recently has the filtering process been studied in detail and also modelled, so that a detailed picture of the water currents has been obtained. In the solitary and most of the colonial choanoflagellates, the water flows freely around the cells, but in some forms, the cells are arranged in an open meshwork through which the water can be pumped. In the sponges, the choanocytes are located in choanocyte chambers (or choanocyte areas) with separate incurrent and excurrent canals/pores located in a larger body, which enables a fixed pattern of water currents through the collar complexes. Previous theories for the origin of sponges show evolutionary stages with choanocyte chambers without any opening or with only one opening, which makes separation of incurrent and excurrent impossible, and such stages must have been unable to feed. Therefore a new theory is proposed, which shows a continuous evolutionary lineage in which all stages are able to feed by means of the collar complexes.     

Canal systems and choanocyte chambers in freshwater sponges (Porifera,Spongillidae)

Summary The spongillid species Spongilla lacustris and Ephydatia fluviatilis possess choanocyte chambers of the classical eurypylous type. They are surrounded by the mesenchymal tissue and connected to the incurrent canal system by prosopyles and to the excurrent canal system by wide apopyles. Each apopyle is sealed against spaces between the basal choanocyte collar parts by a ring of uniflagellated cone cells. The functional aspects of the choanocyte chamber and canal structure are discussed.Dedicated to Prof. Dr. K.E. Wohlfarth-Bottermann, Bonn, in honor of his 65th birthday     

Feeding in a calcareous sponge: particle uptake by pseudopodia

Sponges are considered to be filter feeders like their nearest protistan relatives, the choanoflagellates. Specialized "sieve" cells (choanocytes) have an apical collar of tightly spaced, rodlike microvilli that surround a long flagellum. The beat of the flagellum is believed to draw water through this collar, but how particles caught on the collar are brought to the cell surface is unknown. We have studied the interactions that occur between choanocytes and introduced particles in the large feeding chambers of a syconoid calcareous sponge. Of all particles, only 0.1-microm latex microspheres adhered to the collar microvilli in large numbers, but these were even more numerous on the choanocyte surface. Few large particles (0.5- and 1.0-microm beads and bacteria) contacted the collar microvilli; most were phagocytosed by lamellipodia at the lateral or apical cell surface, and clumps of particles were engulfed by pseudopodial extensions several micrometers from the cell surface. Although extensions of the choanocyte apical surface up to 16 microm long were found, most were 4 microm long, twice the height of the collar microvilli. These observations offer a different view of particle uptake in sponges, and suggest that, at least in syconoid sponges, uptake of particles is less dependent on the strictly sieving function of the collar microvilli.     

Accuracy,Precision, and Observation Rates of Global Positioning System Telemetry Collars

ABSTRACT We addressed concerns regarding performance of various Global Positioning System (GPS) collar configurations for describing habitat use by Rocky Mountain elk (Cervus elaphus) in rugged, forested terrain. We tested 8 GPS collars (Lotek Wireless, Newmarket, ON, Canada) in 4 different model and equipment configurations at 2 reference points (an open hilltop and a forested ravine) to determine habitat-specific differences in performance among collar configurations. We then placed individual collars at 60 additional points that were stratified randomly among 4 canopy-cover classes and 3 classes of available sky. All collars exhibited a locational bias of 4 m horizontally west and of 10 m vertically below a reference standard established by position-averaging with a handheld receiver (Garmin 12MAP) calibrated at National Geodetic Survey benchmarks. The GPS collar models that were programmed for longer satellite-acquisition times provided greater location precision than models that had been programmed for short acquisition times to preserve battery power. Canopy cover and available sky had a greater effect on collar location precision and observation rates than slope, slope position, aspect, conifer basal area, tree height, canopy depth, or elevation. Researchers should test collars at known reference points to confirm that location precision and rates of observation are adequate for their particular study objectives. Manufacturers of GPS collars should inform clients of their programming criteria for acquisition time so that customers can make informed decisions regarding trade-offs between precision of locations, data quantity, and battery life.     

Effects of Global Positioning System Collar Weight on Zebra Behavior and Location Error

ABSTRACT Global Positioning System (GPS) collars are increasingly being used to study fine-scale patterns of animal behavior. Previous studies on GPS collars have tried to determine the causes of location error without attempting to investigate whether the accuracy of fixes provides a correspondingly accurate measure of the animal's natural behavior. When comparing 2 types of GPS collar, we found a significant effect of collar weight and fit on the rate of travel of plains zebra (Equus burchelli antiquorum) females in the Makgadikgadi, Botswana. Although both types of collar were well within accepted norms of collar weight, the slightly heavier collars (0.6% of total body mass [TBM]) reduced rate of travel by >50% when foraging compared with the collar that was 0.4% of TBM. Collar effect was activity specific, particularly interfering with grazing behavior; the effect was less noticeable when zebras crossed larger interpatch distances. We highlight that small differences in collar weight or fit can affect specific behaviors, limiting the extrapolation of fine-scaled GPS data. This has important implications for wildlife biologists, who hitherto have assumed that collars within accepted weight limits have little or no effect on animal movement parameters.     

Fix Success and Accuracy of Global Positioning System Collars in Old-Growth Temperate Coniferous Forests

Abstract: Global Positioning System (GPS) telemetry is used extensively to study animal distribution and resource selection patterns but is susceptible to biases resulting from data omission and spatial inaccuracies. These data errors may cause misinterpretation of wildlife habitat selection or spatial use patterns. We used both stationary test collars and collared free-ranging American black bears (Ursus americanus) to quantify systemic data loss and location error of GPS telemetry in mountainous, old-growth temperate forests of Olympic National Park, Washington, USA. We developed predictive models of environmental factors that influence the probability of obtaining GPS locations and evaluated the ability of weighting factors derived from these models to mitigate data omission biases from collared bears. We also examined the effects of microhabitat on collar fix success rate and examined collar accuracy as related to elevation changes between successive fixes. The probability of collars successfully obtaining location fixes was positively associated with elevation and unobstructed satellite view and was negatively affected by the interaction of overstory canopy and satellite view. Test collars were 33% more successful at acquiring fixes than those on bears. Fix success rates of collared bears varied seasonally and diurnally. Application of weighting factors to individual collared bear fixes recouped only 6% of lost data and failed to reduce seasonal or diurnal variation in fix success, suggesting that variables not included in our model contributed to data loss. Test collars placed to mimic bear bedding sites received 16% fewer fixes than randomly placed collars, indicating that microhabitat selection may contribute to data loss for wildlife equipped with GPS collars. Horizontal collar errors of >800 m occurred when elevation changes between successive fixes were >400 m. We conclude that significant limitations remain in accounting for data loss and error inherent in using GPS telemetry in coniferous forest ecosystems and that, at present, resource selection patterns of large mammals derived from GPS telemetry should be interpreted cautiously.     

STRUCTURE AND FORMATION OF THE COLLAR IN CHOANOCYTES OF TETILLA SERICA (LEBWOHL), DEMOSPONGE

There are two types of collar in the choanocytes of adult Tetilla serica : one type is a continuous cytoplasmic tube and the other consists of discontinuous microvilli. The former is found in the small flagellated chamber and is considered to belong to a young choanocyte in the process of differertiation. To confirm this idea, very young choanocytes which are about to differentiate the collar were examined during embryogenesis.
The youngest choanocytes are noticed forming aggregations of small cells in 3-day larvae. Around the flagellum in each choanocyte, there is a depression which will become wider. At first, the collar is observed as a ring of cytoplasm; next this extends outward and becomes thinner, and finally it divides into microvilli. The microvillous collar is formed by the opening of vesicles and fusion of their membranes. These vesicles are considered to be derived from the Golgi complex. The process of collar formation through fusion of vesicles is discussed.     

A latex-sponge collar for partial-body plethysmography using anesthetized rodents

A latex-sponge sandwich collar was used for sealing rodents in a partial body plethsymograph. This collar formed a leak-free seal between an aerosol inhalation exposure unit and the plethysmograph. Respiratory frequency was compared in anesthetized rats and hamsters using this collar and other types of collars. The newly designed collars did not hamper breathing patterns and reduced the incidence of deaths observed using collars of another design.     

Grizzly Bear Behavior and Global Positioning System Collar Fix Rates

Abstract: Animal locations collected by Global Positioning System (GPS) collars will represent a biased sample of the sites an animal used if some position fixes fail and if those missed locations do not occur randomly. Probability of a GPS receiver obtaining a position fix is known to decline as canopy cover increases, but the impact of forest canopy cover was insufficient to account for the low fix rates we observed for GPS collars on grizzly bears (Ursus arctos). We tested the hypothesis that GPS fix rates were related to the interaction between animal activity (active vs. resting) and canopy cover by evaluating the following predictions: 1) grizzly bear activity should follow a circadian pattern similar to the circadian fix-rate pattern, 2) grizzly bear use of canopy cover should follow a circadian pattern similar to the circadian fix rates, 3) grizzly bear activity should be related to canopy cover (i.e., bears should rest in areas with relatively high canopy covers and feed and move in relatively open areas), and 4) collar orientation and canopy cover should interact to affect the fix rates of test collars. The GPS fix rates traced a bimodal circadian pattern that was directly related to the circadian pattern of grizzly bear activity. Fix rates declined when bears were more likely to be using denser cover, and fix rates of test collars demonstrated that collar orientation interacted with canopy cover, such that fix rates declined much more with increasing canopy cover when the collar was on its side than when the collar was upright. We concluded that inferences made about grizzly bear microhabitat use, based on GPS locations, will underrepresent high canopy cover sites, especially when grizzly bears are resting there. (JOURNAL OF WILDLIFE MANAGEMENT 72(3):596–602; 2008)     

Comparative study of the choanosome of Porifera: 1. The Homoscleromorpha

The choanoderm and pinacoderm of representatives of the two families of Homoscleromorpha sponges, the Oscarellidae and Plakinidae, have been examined by transmission and scanning electron microscopy. Different fixative procedures have shown the dramatic influence of fixation conditions on the morphology of choanocytes. These two families of sponges have the following morphological features in common: flagellated endopinacocytes with short apical microvilli and basal pseudopods; the presence of a very thin and dense sheet of matrix material which limits the mesohyl. There are, however, only minor differences in the flagellar morphology, granule content, and anchoring system of their choanocytes. Two findings are of particular interest: (1) the presence of glycocalyx bridges between the microvilli of the choanocyte collar; and (2) the discovery of a new cell type, the apopylar cell, which has a morphology intermediate between that of pinacocytes and choanocytes. The apopylar cells limit the apopylar opening of the choanocyte chamber and indicate the transition between choanoderm and pinacoderm.     

Oogenesis and larval development of Scypha ciliata (Porifera,Calcarea)

Summary Scypha ciliata is a syconoid sponge. Its oocytes differentiate from choanocytes located near the apopyle of a flagellated chamber, and initially they remain in that location, in a trophic complex with neighbouring choanocytes. When this first growth phase is completed, the oocyte migrates to the periphery of the sponge. There it undergoes a second growth phase, in which it phagocytizes choanocytes and mesenchyme cells.Fertilization of the mature egg is assisted by a converted choanocyte, the sperm carrier cell. This cell penetrates the oocyte and transfers to it the sperm contained in a carriercell vacuole. No meiotic events have yet been observed.Cleavage is asynchronous, with holoblastic, approximately equal divisions. After the first cleavage steps the blastomeres often contain multiple nuclei. The single-layered blastoderm of the stomoblastula consists of many micromeres with flagella that project into the blastocoel, a few macromeres and four cruciform cells. There is no development of a follicle epithelium.The stomoblastula develops into the amphiblastula by inversion; with the assistance of the maternal choanocyte epithelium, the hollow sphere turns inside out, simultaneously moving out of the mesoderm and into the lumen of the adjacent flagellated chamber. In this process, the blastocoel of the stomoblastula is lost. The flagellated cells that form the wall of the amphiblastula now have their flagella extending outward; the amphiblastula also comprises four cruciform cells, macrogranular and agranular cells. The larval cavity of the amphiblastula is a newly formed structure.Abbreviations AB amphiblastula - AP apopyle - BC blastocoel - aC agranular cell - maC macrogranular cell - miC microgranular cell - CB crystalline body - CC central cavity - Ch choanocyte - fCh flat choanocyte - gCh granulate choanocyte - CM cell membrane - Co collar of choanocyte - CrC cruciform cell - DM dense material - EM electron micrograph - F flagellum - FC flagellated cell - FCm flagellated chamber - FL free larva - FV food vacuole - IR interior region - LC larval cavity - M mesenchyme - Ma macromere - MC mesenchyme cell - Mi micromere - N nucleus - Nu nucleolus - O opening - OC oocyte - P psudopodium - PC pinacocyte - PhM phase-contrast micrograph - Po pore - PP prosopyle - S sperm - SB stomoblastula - SC segmentation cavity - SCC sperm-carrier cell - SV sperm vacuole - lT large trophocyte - sT small trophocyte - V vacuole - VC vesicular cytoplasm - VM vacuole membrane     

‘Collar propagation’ as an alternative propagation method for rhizomatous miscanthus

The demand for perennial nonfood crops, such as miscanthus, is increasing steadily, as fossil resources are replaced by biomass. However, as the establishment of miscanthus is very expensive, its cultivation area in Europe is still small. The most common propagation method for miscanthus is via rhizomes, the harvesting of which is very labour‐intensive. Seed propagation is promising, but not suitable for sterile genotypes. In this study, a new vegetative propagation method, ‘collar propagation’, was tested in field and controlled environment studies. Collars are built at the junction between rhizome and stem. They can be harvested in a less destructive way than rhizomes by pulling out the stems from winter‐dormant miscanthus plants. One genotype of each of the species M. sacchariflorus, M. × giganteus, M. sinensis in combination with three fragment types (collars, rhizomes, collars + rhizomes) were tested for establishment success and plant performance. The performance (e.g. dry matter yield) of collar‐propagated plants was either better than or not significantly different from rhizome‐propagated plants. Pregrown plantlets transplanted into the field showed no significant differences in establishment success between the fragments within a genotype. When directly planted into the field however, the fragment ‘rhizome+collar’ had a significantly better establishment success than the other two. The winter survival rate of the fragment ‘rhizome+collar’ was 70% for M. sacchariflorus and 75% for M. × giganteus. Emergence success from collar‐derived plants was not affected by harvest date (harvested monthly from November to February). This study showed that miscanthus propagation via collars is feasible and a promising alternative to rhizome propagation, as the multiplication rate of collars is comparable to that of rhizome propagation. Collar propagation is the more suitable method for the tested genotypes of the species M. sachariflorus and M. × giganteus, but not for M. sinensis genotypes, which may be better propagated by seeds.     

Testing VHF/GPS Collar Design and Safety in the Study of Free-Roaming Horses

Effective and safe monitoring techniques are needed by U.S. land managers to understand free-roaming horse behavior and habitat use and to aid in making informed management decisions. Global positioning system (GPS) and very high frequency (VHF) radio collars can be used to provide high spatial and temporal resolution information for detecting free-roaming horse movement. GPS and VHF collars are a common tool used in wildlife management, but have rarely been used for free-roaming horse research and monitoring in the United States. The purpose of this study was to evaluate the design, safety, and detachment device on GPS/VHF collars used to collect free-roaming horse location and movement data. Between 2009 and 2010, 28 domestic and feral horses were marked with commercial and custom designed VHF/GPS collars. Individual horses were evaluated for damage caused by the collar placement, and following initial observations, collar design was modified to reduce the potential for injury. After collar modifications, which included the addition of collar length adjustments to both sides of the collar allowing for better alignment of collar and neck shapes, adding foam padding to the custom collars to replicate the commercial collar foam padding, and repositioning the detachment device to reduce wear along the jowl, we observed little to no evidence of collar wear on horses. Neither custom-built nor commercial collars caused injury to study horses, however, most of the custom-built collars failed to collect data. During the evaluation of collar detachment devices, we had an 89% success rate of collar devices detaching correctly. This study showed that free-roaming horses can be safely marked with GPS and/or VHF collars with minimal risk of injury, and that these collars can be a useful tool for monitoring horses without creating a risk to horse health and wellness.     

COMPARATIVE ANALYSES OF THE DINOFLAGELLATE FLAGELLAR APPARATUS. II. CERATIUM HIRUNDINELLA1

The three-dimensional structure of the flagellar apparatus in the gonyaulacoid dinoflagellate. Ceratium hirundinella var. furcoïdes (Schröder) Hub.-Pest. was determined using serial section electron microscopy. The flagellar apparatus is quite large and consists of several components. The two basal bodies nearly abut at their proximal ends and are separated by an angle of approximately 120° The broad longitudinal microtubular root extends from the cell's left edge of the longitudinal basal body and bends around the sulcal/cingular depression into the cell's left antapical horn. A transverse striated fibrous root is associated with the transverse basal body and a narrow electron dense extension is present along the anterior edge of the transverse basal body. This study revealed severa1 hitherto unreported fibrous components of the flagellar apparatus that link the various microtubular and fibrous components to themselves and to the two striated collars. A large striated fibrous connective links the two striated collars to one another. This fibrous connective is linked to another striated fibrous connective that originates from the longitudinal basal body and lies perpendicular to the longitudinal microtubular root. The readily identifiable and numerous components of the Ceratium flagellar apparatus are comparable to those of other dinoflagellates. The combined presence of well dpveloped striated collars, a striated collar connective, and a basal body angle of approximately 120° indicates that this flagellar apparatus is most like that described for Peridinioid dinoflagellates. Important similarities are also noticeable between this flagellar apparatus and that of Oxyrrhis marina.     

Ontogeny of the collar cord: Neurulation in the hemichordate Saccoglossus kowalevskii

The chordate body plan is characterized by a central notochord, a pharynx perforated by gill pores, and a dorsal central nervous system. Despite progress in recent years, the evolutionary origin of each of theses characters remains controversial. In the case of the nervous system, two contradictory hypotheses exist. In the first, the chordate nervous system is derived directly from a diffuse nerve net; whereas, the second proposes that a centralized nervous system is found in hemichordates and, therefore, predates chordate evolution. Here, we document the ontogeny of the collar cord of the enteropneust Saccoglossus kowalevskii using transmission electron microscopy and 3D‐reconstruction based on completely serially sectioned stages. We demonstrate that the collar cord develops from a middorsal neural plate that is closed in a posterior to anterior direction. Transversely oriented ependymal cells possessing myofilaments mediate this morphogenetic process and surround the remnants of the neural canal in juveniles. A mid‐dorsal glandular complex is present in the collar. The collar cord in juveniles is clearly separated into a dorsal saddle‐like region of somata and a ventral neuropil. We characterize two cell types in the somata region, giant neurons and ependymal cells. Giant neurons connect via a peculiar cell junction that seems to function in intercellular communication. Synaptic junctions containing different vesicle types are present in the neuropil. These findings support the hypotheses that the collar cord constitutes a centralized element of the nervous system and that the morphogenetic process in the ontogeny of the collar cord is homologous to neurulation in chordates. Moreover, we suggest that these similarities are indicative of a close phylogenetic relationship between enteropneusts and chordates. J. Morphol., 2010. ©2010 Wiley‐Liss, Inc.     

The central cells of sponges

Summary Twenty-one species of Porifera have been surveyed by light microscopy for the presence, form, and relative abundance of a little known cell type known as central cells. They are found to be present in fifteen of these species and occur in six morphologically recognizable forms. Their functional roles are reinterpreted in the light of new distributional and abundance data. The central cells of the siliceous ceractinomorph demosponges are common and intimately associated with the choanocyte population. They probably play an important role in control of water currents within individual choanocyte chambers to mediate cleaning of the outer chamber surfaces. The central cells of keratosan and tetractinomorph demosponges appear to represent stages in egestive processes of wandering mesenchyme cells.This investigation was supported by an operating grant, No. A9554, from the National Research Council of Canada