Structure and cuticle formation of the reticulated carapace of the ostracode Bicornucythere bisanensis

Electron microscopic examination revealed fine structures of the adult carapace of Bicornucythere bisanensis , which consists mainly of epicutiele, procuticle, membranous layer, epidermal cells and subdermal cells. Piled membrane structure is recognized as an organic framework of the procuticle. Newly formed cuticle examined with specimens at various stages of the molt cycle clarified the process of cuticle formation. Analysis of the complicated process of cuticular ridge formation permits morphogenetic discussion of surface features of the ostracode carapace of many species. Cuticle formation in close relation to cell boundaries clearly indicates that each polygon of the carapace reticulation is formed by one epidermal cell. The cell-polygon correspondence suggests that ostracode reticulation is important for elucidation of the phylogenic and ontogenic development of multicellular organisms at cell level.
□ Ostracoda, carapace structure, biomineralization, cuticle formation.     

The structure of the ostracode carapace

The carapace ultrastructure of three Recent [Heterocypris incongruens (Ramdohr), Cypridopsis uidua (Müller), and Conchoecia belgica Müller] and one Cretaceous ostracode species [Cypridea propunctata Sylvester-Bradley] has been studied by means of standard electron microscope techniques. The carapace is shown to be an organic structure divisible into an outer epicuticle, a median exocuticle and an inner endocuticle in the three cypridacean ostracodes examined. In Conchoecia belgica only an outer epicuticle and an inner endocuticle were observed, but this may be due to preservation. The chitin structure of the carapace of calcareous ostracodes is shown to be essentially an interlocking lattice, more coarsely developed in the exocuticle; layered (lamellar) chitin is only present in the selvage and in the connective tissue joining the valves at the hinge. The non-calcareous Conchoecia belgica possesses the layered chitin structure common to other crustaceans and to insects. The relationship of the carapace structure to that present in the decapod and insect exoskeleton is discussed. The technical problems met with during this study are considered in order to outline the difficulties associated with the study of organisms of this size.     

Find of Tremadocian ostracodes in cherts of Kazakhstan

First late Tremadocian (Early Ordovician) ostracodes were found in cherts of the Burubaital Formation near the “Burultas” deposit in southern Kazakhstan. The preservation of carbonate ostracode shellshells in siliceous environment is discussed. The monotypic ostracode assemblage includes the newly described form, Burultalina nikitinae gen. et sp. nov.     

Skeletal morphogenesis in some living collosphaerid Radiolaria

Two major patterns of shell morphogenesis occurring in varying proportions among species were identified by examining skeletons from five species of living collosphaerid, colonial Radiolaria: Collosphaera huxleyi, Acrosphaera cyrtodon, Acrosphaera spinosa, Siphonosphaera tubulosa and Siphonosphaera socialis. Skeletons possessing large open-lattice structures characterized by ellipsoidal to polygonal pores separated by narrow bars are produced by repeated subdivision of large pores into smaller pores. A bridge-like bar grows across the pore and subdivides it into two or more smaller pores. In skeletons with small, nearly circular pores and occasionally elongated tubular rims, the pores appear to be developed by rim thickening. Silica is deposited at the perimeter of the pore, thus decreasing its diameter and increasing the bar width between pores. In some species exhibiting intermediate types of skeletal morphology, there appears to be varying amounts of both bridge growth and rim growth. These data are used to explore possible phylogenetic pathways for a variety of collosphaerid species and to elucidate the processes of skeletal deposition in Radiolaria.     

Structure and function of vanadium compounds in living organisms

Vanadium has been recognized as a metal of biological importance only recently. In this mini-review, its main functions uncovered during the past few years are addressed. These encompass (i) the regulation of phosphate metabolizing enzymes (which is exemplified for the inhibition of ribonucleases by vanadate), (ii) the halogenation of organic compounds by vanadate-dependent non-heme peroxidases from seaweeds, (iii) the reductive protonation of nitrogen (nitrogen fixation) by alternative, i.e. vanadium-containing, nitrogenases from N₂-fixing bacteria, (iv) vanadium sequestering by sea squirts (ascidians), and (v) amavadine, a low molecular weight complex of V(IV) accumulated in the fly agaric and related toadstools. The function of vanadium, while still illusive in ascidians and toadstools, begins to be understood in vanadium-enzyme interaction. Investigations into the structure and function of model compounds play an increasingly important role in elucidating the biological significance of vanadium.     

Test ultrastructure of some living benthic foraminifers

Development of test ultrastructure through ontogeny is traced in four living species from the foraminiferal families Bolivinitidae and Caucasinidae. Test construction conforms in general to the monolomellar concept, but differs in having double pore membranes between consecutive calcitic lamellae and in maintaining a nearly consistent wall thickness through out ontogeny. The test wall observed by scanning electron microscopy consists of an inner, spongy organic lining, a basal membrane, the calcitic wall with a surface veneer, and an outer membrane. Calcitic two-layered lamellae enclosed by organic membranes are added to the test with each new chamber. Single or multiple organic membrane complexes seal completely the majority of the pores and preclude the free exchange of protoplasm to the test surface.     

Janischewskyidae n. fam. and the duplicature of Palaeozoic ostracodes

A duplicature with a very broad inner calcified lamella is revealed in representatives of the genus Janischewskya Batalina 1926. The presence of this structure necessitates the elimination of Janischewskya from the family Hollinidae. A new family of unknown ordinal disposition – Janischewskyidae – is erected. At present, a duplicature is ascertained in four groups of Palaeozoic ostracodes – Podocopina, Parapodocopina, Selebratina-Coryellina , Janischewskyidae. □stracoda, Janischewskyidae, inner calcified la-mella, Carboniferous.
     

Turtle carapace anomalies: the roles of genetic diversity and environment

Background

Phenotypic anomalies are common in wild populations and multiple genetic, biotic and abiotic factors might contribute to their formation. Turtles are excellent models for the study of developmental instability because anomalies are easily detected in the form of malformations, additions, or reductions in the number of scutes or scales.

Methodology/Principal Findings

In this study, we integrated field observations, manipulative experiments, and climatic and genetic approaches to investigate the origin of carapace scute anomalies across Iberian populations of the European pond turtle, Emys orbicularis. The proportion of anomalous individuals varied from 3% to 69% in local populations, with increasing frequency of anomalies in northern regions. We found no significant effect of climatic and soil moisture, or climatic temperature on the occurrence of anomalies. However, lower genetic diversity and inbreeding were good predictors of the prevalence of scute anomalies among populations. Both decreasing genetic diversity and increasing proportion of anomalous individuals in northern parts of the Iberian distribution may be linked to recolonization events from the Southern Pleistocene refugium.

Conclusions/Significance

Overall, our results suggest that developmental instability in turtle carapace formation might be caused, at least in part, by genetic factors, although the influence of environmental factors affecting the developmental stability of turtle carapace cannot be ruled out. Further studies of the effects of environmental factors, pollutants and heritability of anomalies would be useful to better understand the complex origin of anomalies in natural populations.     

Extracellular secretion of phenolic substances from living brown algae

The chemical structures of ultraviolet (UV)- absorbing substances secreted from the healthy living brown algae, Eisenia bicyclis and Ecklonia kurome, were demonstrated. The living activity of algal cells was critically examined using a confocal laser-scanning microscope after incorporation of fluorescein diacetate (FDA) into the cells. Using thin-layer chromatography (TLC), reversed-phase three-dimensional high-performance liquid chromatography (RP-3D-HPLC) and gas chromatography-mass spectrometry (GC-MS), it was found that the UV-absorbing substances (λ_max 265–270 nm) secreted from the living brown algae mainly consisted of the three monomeric bromophenols, 2,4-dibromophenol, 2,4,6-tribromophenol and dibromo-iodophenol, but not phloroglucinol or phlorotannins. The other minor compounds detected in the secretions were as follows: benzothiazole, fatty acids (14:1, 16:0 and 18:0 acids), franesol, 3-hydroxy-2,4,4-trimethylpentyl 2-methylpropanoate and squalene. Exudation of phloroglucinol and phlorotannins was ascertained to take place only after the cell death of these algae. These results indicate that, whilst the algae are alive, polymeric phlorotannins are strictly kept within the algal body, and only monomeric bromophenols are secreted into the seawater medium.     

The intracellular transport and secretion of calumenin-1/2 in living cells

Calumenin isoforms 1 and 2 (calu-1/2), encoded by the CALU gene, belong to the CREC protein family. Calu-1/2 proteins are secreted into the extracellular space, but the secretory process and regulatory mechanism are largely unknown. Here, using a time-lapse imaging system, we visualized the intracellular transport and secretory process of calu-1/2-EGFP after their translocation into the ER lumen. Interestingly, we observed that an abundance of calu-1/2-EGFP accumulated in cellular processes before being released into the extracellular space, while only part of calu-1/2-EGFP proteins were secreted directly after attaching to the cell periphery. Moreover, we found the secretion of calu-1/2-EGFP required microtubule integrity, and that calu-1/2-EGFP-containing vesicles were transported by the motor proteins Kif5b and cytoplasmic dynein. Finally, we determined the export signal of calu-1/2-EGFP (amino acid positions 20-46) and provided evidence that the asparagine at site 131 was indispensable for calu-1/2-EGFP stabilization. Taken together, we provide a detailed picture of the intracellular transport of calu-1/2-EGFP, which facilitates our understanding of the secretory mechanism of calu-1/2.     

The muscle scar in cavellinids and its importance for the phylogeny of platycope ostracodes

The adductor muscle scars of cavellinids from the Visean (in one case from the Upper Devonian) of the European part of the RSFSR have revealed a significant diversity in their morphology. Aggregate, multiserial with up to five series, bi- to triserial and nearly biserial muscle scars existed simultaneously. The verticaliserial muscle scars were initiated by a biserial larval group which in the course of evolution may have passed, due to neoteny, into the adult stage. Together with Cytherella sp. from Syria, the new data show that the genus Cytherella appeared not in Triassic nor Jurassic time, hut originated among cavellinids possibly in the Early Carboniferous. A new genus is Borovitchella , with the type species B. egorovi n. sp.     

Functional morphology and mode of life of Palaeozoic leiocope ostracodes

Leiocopes differ from all other major ostracode groups (e.g. Palaeocopa. Binodicopa, Podocopa, Myodocopa) mainly in having a simple rounded/convex shape and a strong (dorsal and/or ventral) asymmetry. These morphological features are highly conservative in leiocopes through the Palaeozoic (lower Ordovician-middle Devonian). Functional morphology and facies distribution suggest that typical 'dome-like' leiocopes (e.g. aparchitids) may have lived off the bottom in relatively deep water environments, contrasting with most (inferred) benthic Palaeozoic ostracodes (e.g. heavily calcified dimorphic palaeocopes). Reversely, smaller sized and both dorsally and ventrally asymmetrical leiocopes (e.g. Ordovician jaanussoniids) are considered as possible benthic dwellers with reduced mobility on or within the substratum. □ Ostracodes, Leiocopa, Aparchitidae, Jaanussoniidae. Palaeozoic. Ordovician, Silurian, Devonian, asymmetry, functional morphology.     

Hepatocyte growth factor is crucial for development of the carapace in turtles

Turtles are characterized by their shell, composed of a dorsal carapace and a ventral plastron. The carapace first appears as the turtle-specific carapacial ridge (CR) on the lateral aspect of the embryonic flank. Accompanying the acquisition of the shell, unlike in other amniotes, hypaxial muscles in turtle embryos appear as thin threads of fibrous tissue. To understand carapacial evolution from the perspective of muscle development, we compared the development of the muscle plate, the anlage of hypaxial muscles, between the Chinese soft-shelled turtle, Pelodiscus sinensis, and chicken embryos. We found that the ventrolateral lip (VLL) of the thoracic dermomyotome of P. sinensis delaminates early and produces sparse muscle plate in the lateral body wall. Expression patterns of the regulatory genes for myotome differentiation, such as Myf5, myogenin, Pax3, and Pax7 have been conserved among amniotes, including turtles. However, in P. sinensis embryos, the gene hepatocyte growth factor (HGF), encoding a regulatory factor for delamination of the dermomyotomal VLL, was uniquely expressed in sclerotome and the lateral body wall at the interlimb level. Implantation of COS-7 cells expressing a HGF antagonist into the turtle embryo inhibited CR formation. We conclude that the de novo expression of HGF in the turtle mesoderm would have played an innovative role resulting in the acquisition of the turtle-specific body plan.     

Ultrastructure of the carapace margin in the Ostracoda (Arthropoda: Crustacea)

In podocopid ostracods, the “hingement”, with teeth/sockets or crenulations, is developed along the attached dorsal margin of the carapace. Dual calcified lamellae, called “duplicatures” are also developed along the free margin. However, the terminology of these marginal areas is often confused by many ostracodologists because of their differing interpretations and viewpoints. In this study, the marginal areas of the carapace were observed in detail, using the transmission (TEM) and scanning (SEM) electron microscopes. Consequently, the terminology of the marginal areas has been revised. The homology between attached and free margins and the structural differences of duplicatures, for some taxa are also discussed.