Ultrastructure of the coxal gland of the horseshoe crab Limulus polyphemus: Evidence for ultrafiltration and osmoregulation

Electron microscopic examination of the paired coxal glands of the horseshoe crab Limulus polyphemus, focusing on urinary and vascular channels, shows six morphologically distinct regions. Each of four nephridial lobes consists of two cortical layers surrounding a medulla. The outer and inner cortexes contain blood vessels separated by a basement membrane from the urinary space lined by podocytes. Podocyte foot processes are applied to the basement membrane, interdigitate with those from other podocytes, and have a filtration slit diaphragm between them. Cortical morphology demonstrates ultrafiltration of blood, a previously undescribed function of the gland, as well as possible endocytic reabsorption of materials by the podocytes. The medulla drains into the stolon connecting the four lobes. These two areas have urinary tubules of cuboidal epithelium featuring microvillous-like apical projections, cytoplasmic vesicles and vacuoles, elaborate lateral interdigitations with septate junctions, and basal invaginations containing numerous mitochondria. These tubules are closely surrounded by blood channels, lined by a basement membrane containing embedded support cells. The medulla and stolon morphology are suggestive of both ion transport and water movement, in keeping with the gland's role in osmoregulation. The stolon empties into the end sac in the base of the most posterior lobe. It is lined by tall epithelium exhibiting apical overlap, blunt projections into the lumen, apparent endocytic vesicles, and basal plasma membrane infoldings with mitochondria. The end sac drains into the conducting nephric duct, the proximal end of which is lined by a cuticle. J. Morphol. 234:233–252, 1997. © 1997 Wiley-Liss, Inc.     

Evolution of adhesive mechanisms in cribellar spider prey capture thread: evidence for van der Waals and hygroscopic forces

Sticky prey capture threads are produced by many members of the spider infraorder Araneomorphae. Cribellar threads are plesiomorphic for this clade, and viscous threads are apomorphic. The outer surface of cribellar thread is formed of thousands of fine, looped fibrils. Basal araneomorphs produce non-noded cribellar fibrils, whereas more derived members produce noded fibrils. Cribellar fibrils snag and hold rough surfaces, but other forces are required to explain their adherence to smooth surfaces. Threads of Hypochilus pococki (Hypochilidae) formed of non-noded fibrils held to a smooth plastic surface with the same force under low and high humidities. In contrast, threads of Hyptiotes cavatus and Uloborus glomosus (Uloboridae) formed of noded fibrils held with greater force to the same surface at intermediate and high humidities. This supports the hypothesis that van der Waals forces allow non-noded cribellar fibrils to adhere to smooth surfaces, whereas noded fibrils, owing to the hydrophilic properties of their nodes, add hygroscopic forces at intermediate and high humidities. Thus, there appear to have been two major events in the evolution of adhesive mechanisms in spider prey capture thread: the addition of hydrophilic nodes to the fibrils of cribellar threads and the replacement of cribellar fibrils by viscous material and glycoprotein glue.  © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 77 , 1–8.     

Direct rhizogenesis, in vitro stolon proliferation and high-throughput regeneration of plantlets in Glycyrrhiza glabra

Direct rhizogenesis from leaf explants and establishment of an in vitro stolon culture system and subsequent plant regeneration for Glycyrrhiza glabra have been described. MS liquid medium supplemented with 0.01 mg l^?1 of NAA was most effective for stolon proliferation. Extensive proliferation of stolon and shoot regeneration was achieved on medium containing 3 % sucrose with 0.01 mg l^?1 NAA. Stolons with nodes showing growth was transferred under light for plantlet regeneration in the same medium. This paper is the first report in G. glabra describing a complete regeneration procedure via in vitro stolon proliferation along with quantitative data for glycyrrhizin and genetic fidelity of plant regenerated in vitro there from. In vitro stolon proliferation described here would be an efficient way for regeneration of plants for functional genomics studies and better understanding of glycyrrhizin (GA) metabolism.     

The Fine Structure of Rhynchocystis pilosa (Sporozoa, Eugregarinida)

SYNOPSIS. The trophozoite of Rhynchocystis pilosa obtained from the seminal vesicles of the earthworm Lumbricus terrestris was studied by light and electron microscopy. The trophozoite's cortical organization is particularly interesting because of its unusual evaginations and associated fibrillar structures. The pellicle is formed by 2 concentric membranes elevated into 60–70 alternating primary and secondary ridges extending posteriad. Numerous long ‘hairs’ or cytopilia originate along the primary ridges and each contains a system of fibrils originating from an underlying longitudinal myoneme. Longitudinal rows of pores lie between adjacent pollicular ridges. Three systems of fibrils lie in the cortex of the trophozoite. A longitudinal myoneme consisting of 12–18 fibrils lies below each primary pellicular ridge. Circular myonemes lie below the pellicle in a parallel array along the length of the organism. Each myoneme consists of 4–8 fibrils structurally similar to those of the longitudinal myonemes. Pairs of fine filaments also lie in the inner pellicular membrane along the apex of each ridge. The trophozoite's anterior end is modified as an attachment organelle consisting of 30–35 delicate pellicular folds which originate at the base of an anterior papilla. The folds extend approximately 15 μ posteriad where they become continuous with the primary pellicular ridges. The nucleus lies in the cytoplasm near the posterior level of the attachment organelle and is surrounded by a double membrane perforated by numerous pores. The cytoplasm contains numerous small vesicles which may be found in dense aggregations. These aggregations often occur in proximity to Golgi complexes and certain membrane-bound bodies. Mitochondria are abundant in the cytoplasm as are large, ovoid paraglycogen bodies. Occasionally layers of granular membranes are arranged parallel to the surface of the paraglycogen bodies but also occur thruout the cytoplasm.     

The dormant buds of Rhabdopleura compacta (Hemichordata)

Rhabdopleura has an overwintering stage that consists of two layers of cells surrounding a central yolk mass. This cellular part is surrounded by a thick electron dense capsule which is secreted by the bud itself. The capsule is probably impervious and protective to its contents. Blood vessels join the buds to the zooids of the colony. They form the probable route of transfer of yolk from the zooids to the dormant bud. The capsule of the dormant bud has some structural features in common with the black stolon of the adult zooids. The black stolon is probably formed in a manner similar to that which made the fusellar fabric of the periderm of fossil graptolities.     

慈姑匍匐茎的起源和球茎的膨大研究

匍匐茎的发生一般见于主茎倒二或倒三叶原基的叶腋部位。在匍匐茎发生区域的主茎一侧,匍匐茎原始细胞的基部形成壳状区;壳状区的形成对匍匐茎原基的外凸起一定作用。匍匐茎无居间分生组织;它的伸长依靠顶端分生组织细胞的横向分裂,使轴向细胞数目增多,并使细胞的轴向延伸。球茎的膨大是通过匍匐茎第8—10节基本分生组织的细胞有丝分裂,增加细胞数目,然后细胞体积的扩大来实现的。球茎中的淀粉一般为单粒淀粉;匍匐茎中的淀粉由单粒和复合两种淀粉粒组成。     

Transgenic zoysiagrass (Zoysia japonica) plants obtained by Agrobacterium-mediated transformation

Zoysiagrass (Zoysia japonica Steud.) is an important turfgrass that spreads by stolons and rhizomes. By exploring the potential of direct shoot formation from stolons, we developed a straightforward and efficient transformation protocol without callus induction and propagation. Sterilized stolon nodes were infected and co-cultivated with Agrobacterium tumefaciens harboring pCAMBIA vectors. Hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin was used as the selection agent. Both green and albino shoots were directly regenerated from the infected stolon nodes 4–5 weeks after hygromycin selection. Greenhouse-grown plants were obtained 10–12 weeks after Agrobacterium-mediated transformation. Based on the number of transgenic plants obtained and the number of stolon nodes infected, a transformation frequency of 6.8% was achieved. Stable integration of the transgenes in the plant genome was demonstrated by PCR and Southern blot hybridization analyses. Expression of the transgenes was confirmed by RT-PCR analysis and GUS staining. The new transformation system opens up new opportunities for the functional characterization of genes and promoters and the development of novel germplasm in zoysiagrass.     

Effects of node position on regeneration of stolon fragments in congeneric invasive and native Alternanthera species in China

Vegetative regeneration plays an important role in the adaptation of clonal plants in frequently disturbed habitats, but few studies have compared vegetative regeneration capacity of invasive clonal plants with that of their native congeners. Vegetative regeneration capacity from shoot nodes can also be affected by the position of the nodes, but this appears little studied. We conducted a greenhouse experiment with Alternanthera philoxeroides, a highly invasive species in China, and its native congener A. sessilis to test the difference in vegetative regeneration capacity of stolon nodes at five different positions (i.e. 3rd, 4th, 5th, 6th and 7th node starting from the apex of the stolon). At the end of the experiment, we counted and harvested all regenerated plants and determined their biomass and allocation. Both species could successfully regenerate from stolon fragments and node position significantly affected regeneration rate and subsequent growth. However, the vegetative regeneration capacity of A. philoxeroides was not higher than that of A. sessilis. These results suggest that vegetative regeneration from stolon fragments may not be a trait that can explain the invasiveness of A. philoxeroides.     

Quantitative measures of gastrovascular flow in octocorals and hydroids: toward a comparative biology of transport systems in cnidarians

Using microscopy, the gastrovascular systems of four hydroids (Eirene viridula, Cordylophora lacustris, Hydractinia symbiolongicarpus, and Podocoryna carnea) and two distantly related alcyonacean octocorals (Acrossota amboinensis and Sarcothelia sp.) were examined and compared within a phylogenetic framework. Despite a range of stolon widths (means 53–160 μm), the hydroid species exhibited similar patterns of gastrovascular flow: sequentially bidirectional flow in the stolons, driven by myoepithelial contractions emanating from the center of the colony. Unlike the hydroids, the gastrovascular system of A. amboinensis (mean stolon widths for 5 colonies, 0.57–1.21 mm) exhibited simultaneously bidirectional flow with incomplete, medial baffles (width 4–20 μm) separating the flow. Baffles visualized with transmission electron microscopy consisted of endoderm, mesoglea, and occasionally another layer of tissue. Mean flow rates of the gastrovascular fluid for seven stolons ranged from 125 to 275 μm s^?1, with maximum rates of 225–700 μm s^?1. In Sarcothelia sp., stolons were of comparable width (means for 13 colonies 0.55–1.4 mm) to those of A. amboinensis. These stolons, however, were divided by several partitions (width 8–25 μm), both complete and incomplete, which were spaced every 100.5±5.1 μm (mean±SE; range 27.1–283.7 μm) and appeared structurally similar to baffles. In lanes defined by these partitions, ciliary motion was visible in image sequences, and flow was unidirectional. Within a single stolon, flow moved in different directions in different lanes and changed direction by moving from lane to lane via occasional spaces between the partitions. Mean flow rates for 30 stolons ranged from 75 to 475 μm s^?1, with maximum rates of 85–775 μm s^?1. For both octocorals, flow rates of the gastrovascular fluid were not correlated with the width of the stolon lumen. While octocoral gastrovascular systems probably exhibit differences based on phylogenetic affinities, in all species studied thus far, gastrovascular flow is entirely driven by cilia, in contrast to the hydroid taxa.     

INTERNAL ARCHITECTURE OF MIOCENE PSEUDOTABERINA AND ITS RELATION TO CARIBBEAN ARCHAIASINS

Abstract:  Well-preserved, isolated specimens of Pseudotaberina were found at three localities in Indonesia, thus enabling the study of the relationships between this genus and Caribbean Archaiasinae. The internal architecture of the chambers of Pseudotaberina consists of four layers of stolon planes, each of which is divided by superposed septula into chamberlets. The chamberlets are connected to the next chambers by foramina opposing the septula. The stolons are Y-shaped. All other Archaiasinae have radial stolon axes combined with an initial involute growth stage, whereas the Soritinae have cross-wise oblique stolon axes, but with evolute growth in the early stages. Pseudotaberina shows a combination of the characters of these two subfamilies, confirming the need to study more Tethyan material to elucidate phylogenetic relationships within this group of large benthic foraminifera.     

Ultrastructure of gill bars of Branchiostoma lanceolatum with special reference to gill skeleton and blood vessels (Cephalochordata)

Summary The skeletal rods of the gill bars of Branchiostoma have been examined histochemically and electron microscopically. The rods are composed of 15-nm thick filaments which are interconnected by 10-nm thick and 15-nm long cross-linkages. The filaments appear to consist of structural proteins that are totally different from the proteins of the collagen fibrils. The cross-linkages between the filaments consist of acid mucopolysaccharides, and probably include chondroitin sulphate. The fine structure of blood vessels in the gill bars has been studied in uninjected specimens and in those intravascularly injected with either india ink or horseradish peroxidase. Blood vessels surrounded by basal laminae of delimiting epithelia or simply surrounded by connective tissue material are present in the gill bars. Very characteristic blood cells are normally found in these vessels. Furthermore, when either india ink or horseradish peroxidase is injected via the aorta and the endostylar vessel, irregular slitlike channels can be seen in the connective tissue. These channels are in open connection with the blood vessels and are thus a part of the vascular system.Supported by a grant from the Danish Natural Science Research Council     

SIF, a novel morphogenetic inducer in hydrozoa.

By analogy to processes in angiogenesis (blood vessel formation), the development of the stolonal network in colonial hydrozoa involves stimulation of branching and mutual chemotropic attraction of the growing branches by means of soluble morphogenetic factors. We have identified a glycoconjugate of about 20 kDa, termed SIF (Stolon-Inducing Factor), which induces the formation of stolon branches when applied locally. Micropipettes ejecting SIF mimic the inducing action of stolon tips, the putative sources of SIF. When whole animals are exposed to SIF, stolons sprout not only from the base of the polyps but also from abnormal sites along the entire body, even from the head. In addition, the polyp (hydranth) secretes a chitinous periderm which, in the species under investigation, normally envelops stolons but not hydranths. At high SIF doses the whole hydranth is transformed into stolon tissue. The factor has been isolated from conditioned medium and from butanol extracts of Hydractinia echinata and Podocoryne carnea.     

End-to-end self-assembly of RADA 16-I nanofibrils in aqueous solutions

RADARADARADARADA (RADA 16-I) is a synthetic amphiphilic peptide designed to self-assemble in a controlled way into fibrils and higher ordered structures depending on pH. In this work, we use various techniques to investigate the state of the peptide dispersed in water under dilute conditions at different pH and in the presence of trifluoroacetic acid or hydrochloric acid. We have identified stable RADA 16-I fibrils at pH 2.0–4.5, which have a length of ～200–400 nm and diameter of 10 nm. The fibrils have the characteristic antiparallel β-sheet structure of amyloid fibrils, as measured by circular dichroism and Fourier transform infrared spectrometry. During incubation at pH 2.0–4.5, the fibrils elongate very slowly via an end-to-end fibril-fibril aggregation mechanism, without changing their diameter, and the kinetics of such aggregation depends on pH and anion type. At pH 2.0, we also observed a substantial amount of monomers in the system, which do not participate in the fibril elongation and degrade to fragments. The fibril-fibril elongation kinetics has been simulated using the Smoluchowski kinetic model, population balance equations, and the simulation results are in good agreement with the experimental data. It is also found that the aggregation process is not limited by diffusion but rather is an activated process with energy barrier in the order of 20 kcal/mol.     

Xylem and Phloem Transport of Mineral Nutrients from Solanum tuberosum Roots

The xylem and phloem transport of mineral elements from stemnodal roots to the stem and stolon of growing potato (Solanumtuberosum L. cv. ‘Russet Burbank’) plants was investigated.Adventitious roots, originating from below-ground nodes of thestem of potato seedlings, were exposed to solutions of SrCI₂or MnSO₄. Relative elemental concentrations were measured inthe conductive tissues using energy dispersive X-ray analysis.After a 5 h daylight uptake period, Sr (a Ca-transport analogue)levels were elevated in the stem xylem tissue, but Sr did notincrease in the stem phloem, nor was it present in either ofthe conductive tissues of stolons located 1–2 nodes abovethe treated roots. In contrast, elevated levels of Cl, S, andMn were found in stolon xylem and phloem tissue during the sameperiod. The absence of Sr in the stolon after 5 h suggests thatno xylem flow into the stolon occurred during the uptake periodand, furthermore, phloem flow is responsible for the transportof the Cl, S, and Mn into the stolon. Elevated levels of thesemobile nutrients in the xylem of the stolon were attributedto xylem-to-phloem transfer in the stem or leaves, transportto the stolon in the phloem, and phloem-to-xylem transfer inthe stolon. During a 19 h uptake period, some Sr was observedin the phloem tissue of the stem, demonstrating slow exchangeof Sr with sieve elements or proximal phloem parenchyma andcompanion cells. Key words: Calcium, manganese, X-ray analysis     

Ultrastructure ofThraustochytrium sp. zoospores

Summary The kinetosome-flagellum base ofThraustochytrium sp. has several ultrastructural features that are unique among the fungi studied thus far. Within the lumen formed by the kinetosome fibrils are two kinds of electron opaque areas. Near the proximal end of the kinetosome is an electron opaque granule or core. Distal to this core is an area where electron opaque substances are found concentrated between adjacent triplet fibrils. Proximal to the terminal plate is an electron opaque disc, the basal disc, whith its complex arrangment of fibrous material, is unlike any reported thus far among the fungi. The nine peripheral flagellar fibrils are surrounded by electron opaque material in a region just distal to the terminal plate.Contribution No. 444, Virginia Institute of Marine Science, Gloucester Point, Virginia 23062, U.S.A.Supported in part by National Science Foundation Grant # GA-31014 to Dr. Frank O. Perkins.     

The envelopes of the nervous system of pseudoscorpions and scorpions

The present paper demonstrates that in pseudoscorpions and scorpions a compact collagen sheath consisting of fibrils with a 720 A period embedded in a neutral mucopolysaccharide surrounds the nervous system. The same material makes up the trabeculae running into the nervous elements of the ganglia and nerves. Both structures are separated from the neurons and their fibres by entire glial cells or by their processes. The neural lamella is surrounded by a thin and continuous wall of cells forming a storage-epithelium. Amoebocytes are able to circulate between the collagen fibrils of the neural lamella.     

Immunocytochemistry of the acellular slime mold Physarum polycephalum. III. Distribution of myosin and the actin-modulating protein (fragmin) in sandwiched plasmodia

The acellular slime mold Physarum forms very thin plasmodia when sandwiched between two agar sheets. After extraction with glycerol-containing buffers, suitable objects for immunofluorescence microscopy are obtained, and an analysis of the cytoskeletal and contractile system of Physarum becomes possible. Plasmodia were stained with antibodies against myosin and fragmin, a protein factor involved in actin filament length regulation. The microanatomy and topography of cellular structures containing these proteins were investigated at the light and electron microscopic levels. The patterns obtained with the two antibodies are closely related to those obtained with actin antibody [25]. In both cases the complex system of cytoplasmic fibrils is stained selectively. The fibrils form a more or less regular network in the advancing front zone with the fibrils being interconnected by focal nodes. In the posterior region of the plasmodium, where endoplasmic pathways and protoplasmic veins are differentiated, larger fibrils are detected, running obliquely or longitudinally to the veins. With both antibodies the fluorescent pattern of the fibrils is continuous without indications of periodic interruptions or striations, which would be expected in the case of sarcomere-like subunits. With anti-myosin unstained patches are frequently seen at or close to the nodes of the fibrillar network in the anterior region. The small lobopodia, which are rich in actin, are apparently not stained by the myosin antibody, a result similar to the situation in "ruffling edges? of cultured vertebrate cells. Electron microscopic investigations of antibody-labeled fibrils in embedded and sectioned plasmodia allow the identification of antibody molecules at specific sites along the fibrils with a different distribution pattern for each of the two antibodies.     

Structure of the temporal organ of the Japanese house centipede,Thereuonema hilgendorfi Verhoeff

The ultrastructure of the temporal organ of the Japanese house centipede, Thereuonema hilgendorfi Verhoeff (Chilopoda), has been examined. The temporal organ of this species is known to be a carbon dioxide receptor. It appears externally as a small protuberance with a small opening (5 μm) on its summit. There is a small cuticle-lined cavity beneath the protuberance, and a mushroom-shaped projection protrudes from the base of the cavity into its interior. Below the cavity is an encapsulated, bulb-shaped cellular mass (sensory bulb), which contains about ten receptor cells and 100 supporting cells. Nuclei of both receptor and supporting cells occur basally in the sensory bulb. Each receptor cell has a single dendrite, which gives rise to a pair of sensory cilia. The cilia enter the mushroom-shaped projection, and course along the inner surface of its calyx. The supporting cells have distal processes, which accompany the cilia into the calyx. The surface cuticle of the calyx consists of a fabric of fibrils, but in not layered like the cuticular integument. The fabric is permeable to water. The observed structure is discussed in relation to carbon dioxide reception and in comparison with the structure of olfactory receptors and hygroreceptors.     

Koskinoid perforations in brachiopod shells: function and mode of formation

Clusters of tiny perforations called 'koskinoid' have been interpreted as having accommodated byssus-like fibrils for attachment in Devonian Uncites and the Orthotetacea, which range from the Silurian through the Permian. The mode of formation of the tiny holes has inspired three hypotheses: (1) The fibrils protruded at the commissure and then were surrounded by growing shell. (2) The fibrils protruded from a juvenile foramen at the beak, which then was covered by shell that caused them to separate and migrate. (3) The fibrils penetrated their own shells like pedicles of living brachiopods are known to penetrate calcite. The first two hypotheses assume that the fibrils were produced by a rudimentary pedicle within the shell; the third postulates that the fibrils are diverticula from the outer epithelium of the mantle. The facts that the clusters are not confined to the beak region, but occur scattered over the ventral valve, and that they also penetrate the interara, favor the third theory. All Orthotetacea except Thecospira have the koskinoid perforations; all Davidsoniacea except Morinorhynchus lack them. This suggests that the two genera should trade places in the classification.