Stereom morphogenesis and differentiation during regeneration of adambulacral spines of Asterias rubens (Echinodermata,Asteroida)

Summary The very first mineral deposits appearing in regenerating fractured adambulacral spines of Asterias rubens are minute polyhedrons that cover the surface of fractured trabeculae. Polyhedrons fuse together forming a fold from which a microspine differentiates. Microspines develop into long linear trabeculae which send out lateral processes at regular length intervals. Lateral processes from adjacent trabeculae fuse together, bridging the trabeculae and giving the regenerate the typical meshwork structure of stereom. Most of the regenerate is built up according to this growth pattern which ensures its longitudinal growth. Simultaneously, the initial fascicular stereom of the stub sends out short radial processes which branch into upward and downward directed subprocesses. The latter fuse with their equivalents located above or below, building up longitudinal rows of stereom meshes. These rows then bridge together by additional branched or unbranched lateral processes, so forming a new stereom layer which progressively covers the whole stub. Up to three new layers of stereom are formed in this way at the stub periphery. These become continuous with the stereom layers of the regenerate by fusion of reciprocal subprocesses, so ensuring the continuity between the stub and the regenerate. In both structures the first stage of mineralization results in an open stereom. Stereom thickening occurs in a second stage of mineralization (that is chronologically separated from the formation of the open stereom) and results in the differentiation of the original stereom fabrics (i.e. fascicular stereom). Regeneration of removed spines starts with the formation of a new spine base made of labyrinthic stereom. The development of the latter mostly relies on short branched and unbranched processes which fuse with each other or with predifferentiated meshes. After completion of its base, the regenerating spine lengthens and thickens similarly to the regenerating fractured spines. The diversity of the stereom growth processes observed in the present work may be reduced to the combination of one to three elementary events, viz. the development of long linear processes, of short unbranched processes and of short branched processes. A survey of the literature allows the suggestion that the implementation of these elementary events is sufficient to describe most types of stereom morphogenesis.Senior research assistant NFSR (Belgium)     

Osteoderm morphology and development in the nine-banded armadillo, Dasypus novemcinctus (Mammalia, Xenarthra, Cingulata)

Among modern mammals, armadillos (Xenarthra, Cingulata) are the only group that possesses osteoderms, bony inclusions within the integument. Along the body, osteoderms are organized into five discrete assemblages: the head, pectoral, banded, pelvic, and tail shields. The pectoral, banded, and pelvic shields articulate to form the carapace. We examined osteoderm skeletogenesis in the armadillo Dasypus novemcinctus using serial and whole-mount histochemistry. Compared with the rest of the skeleton, osteoderms have a delayed onset of development. Skeletogenesis begins as condensations of osteoblasts secreting osteoid, localized within the papillary layer of the dermis. Osteoderm formation is asynchronous both within each shield and across the body. The first osteoderms to mineralize are situated within the pectoral shield of the carapace, followed by elements within the banded, head, pelvic, and tail shields. In general, within each shield ossification begins craniomedially and proceeds caudally and laterally, except over the head, where the earliest elements form over the frontal and parietal bones. The absence of cartilage precursors indicates that osteoderms are dermal elements, possibly related to the all-encompassing vertebrate dermal skeleton (exoskeleton). The mode of development of D. novemcinctus osteoderms is unlike that described for squamate osteoderms, which arise via bone metaplasia, and instead is comparable with intramembranously derived elements of the skull.     

²⁶Mg labeling of the sea urchin regenerating spine: Insights into echinoderm biomineralization process

This paper reports the results of the first dynamic labeling experiment with regenerating spines of sea urchins Paracentrotus lividus using the stable isotope ²⁶Mg and NanoSIMS high-resolution isotopic imaging, which provide a direct information about the growth process. Growing spines were labeled twice (for 72 and 24 h, respectively) by increasing the abundance of ²⁶Mg in seawater. The incorporation of ²⁶Mg into the growing spines was subsequently imaged with the NanoSIMS ion microprobe. Stereom trabeculae initially grow as conical micro-spines, which form within less than 1 day. These micro-spines fuse together by lateral outgrowths and form a thin, open meshwork (inner stereom), which is subsequently reinforced by addition of layered thickening deposits (outer stereom). The (longitudinal) growth rate of the inner stereom is ca. 125 μm/day. A single (ca. 1 μm) thickening layer in the stereom trabeculae is deposited during 24 h. The thickening process is contemporaneous with the formation micro-spines and involves both longitudinal trabeculae and transverse bridges to a similar degree. Furthermore, the skeleton-forming cells remain active in the previously formed open stereom for at least 10 days, and do not migrate upwards until the end of the thickening process. The experimental capability presented here provides a new way to obtain detailed information about the skeleton formation of a multitude of marine, calcite producing organisms.     

Microscopical Anatomy of the Cyrtocrinid Cyathidium meteorensis (sive foresti) (Echinodermata,Crinoidea)

Light microscopy and scanning electron microscopy of three specimens of Cyathidium meteorensis (order Cyrtocrinida) revealed some special morphological features. The brachial articulation is provided with a long tendon at the aboral side; the entire articulation surface, including the areas where ligaments attach is built up by labyrinthic stereom. The calycinal ossicle lacks any internal vestiges of a pentameric composition; vertical planes with changing stereom direction lie irrespectively of radial or interradial planes. Gut, ovary and testis are histologically inconspicuous, and the location of the gonads inside the calyx is quite unusual. Both sexes develop outer gonoducts which probably are functionally adapted madreporic canals. The coelomic system differs from that of other crinoids in that a chambered organ is completely lacking. Simultaneously, the aboral nervous subsystem has no aboral nerve centre and simply terminates aborally in the ring-shaped commissure. A glandular axial organ is absent, as are typical sacculi. The data are in accordance with two proposals made previously for Holopus rangii, viz., that the animals can feed raptorially, and that cyrtocrinids probably have evolved by loss of aboral calycinal ossicles of ancestors. In part, our observations differ from those in Cyathidium foresti, so we have chosen to use the species name meteorensis which has been considered a synonym of foresti.     

Skeleton resorption in echinoderms: Regression of pedicellarial stalks inSphaerechinus granularis (Echinoida)

Summary When a globiferous pedicellaria ofSphaerechinus granularis injects its venom, the head autotomizes whereas the stalk remains on the test and enters a regression process with concomitant resorption of its supporting ossicle (i.e. the rod). Scanning electron microscope investigations of the morphological changes undergone by the stereom of resorbing rods show that: (1) resorption proceeds both axially and laterally, and leads to a reduction of approximately 80% of the original length of the rod, (2) secondary growth of new stereom processes occurs concomitantly with resorption but never ensures even a partial regeneration of the rod, and (3) resorption and secondary growth stop before the rod is totally destroyed leaving a static stump that remains in place up to 190 days. Particular resorption figures result from either the axial or the lateral resorption of the rod shaft. These consist chiefly of terraced conical cupules, dense cylinders and concentric lamellae whose walls or edges are typically made of closely piled and/or aligned subprismatic crystallites. Whatever their location along the rod, these crystallites always organize strictly parallel to the rod axis. Whether the crystallites are mosaic blocks composing larger monocrystalline units or discrete monocrystals themselves is for the moment unclear. A growth model, which accounts for the observed resorption figures, is proposed for the shaft of pedicellarial rods. According to this model, the early growth of the shaft would produce elongated, interconnected trabeculae (initial trabeculae) made of densely piled and perfectly aligned crystallites. Thickening and coalescence of adjoining trabeculae would progressively occur by adjunction around the initial trabeculae of successive and concentric layers of similarly arranged crystallites. Coalescent trabeculae would then be cemented together in a perforate stereom layer by the final deposition of larger crystallite layers surrounding the whole shaft periphery. Growth of secondary stereom processes occurs both in the resorbing rod (here the newly formed processes are resorbed soon after they have been produced) and in rods where resorption has stopped. These are always irregular processes that localize near or on the actual sites of resorption. It is suggested these processes result from an uncontrolled activation of the skeleton-forming cells in areas where the concentration of calcium ions increases as a consequence of calcite resorption.     

Fine structure of the dorsal arm plate of Ophiocoma wendti: Evidence for a photoreceptor system (Echinodermata,Ophiuroidea)

Summary Three structures in the dermis of the dorsal arm plate (DAP) of the brittlestar, Ophiocoma wendti, appear to comprise a photoreceptor system. The upper surface of the DAP bears transparent, knob-like, microscopic structures which are expanded peripheral trabeculae (EPT) of the calcite stereom. The EPT are part of the photoreceptor system and can facilitate light transmission through the DAP by decreasing light refraction, reflection and absorption that occur at stereom/stroma interfaces. Bundles of nerve fibres located below the EPT are a second component of the system, and may function as primary photoreceptors. The intensity of light impinging on the putative sensory tissue is regulated by the diurnal activity cycle of chromatophores, the third element of the system. During the day the chromatophores cover the EPT and thereby shade the nerve fibres. At night they retract into inter-trabecular channels, uncovering the EPT and thereby exposing the nerve fibres to transmitted light. Thus, the transparent stereom may play a role in photoreception, in addition to its generally recognized skeletal function. Although ciliated cells that may be sensory are present in the epidermis of Ophiocoma wendti, they do not appear to be photoreceptors. Functional analogues of the brittlestar photoreceptor system in other echinoderms are discussed, emphasizing the relationship between photosensitivity and the transparency of the stereom in several classes of Echinodermata.     

Middle Cambrian cystoid (sensu lato) stem columnals from Bornholm, Denmark

A rich material of echinoderm fragments from two Middle Cambrian stratigraphical levels on Bornholm are preserved due to phosphatization of the original calcitic stereom. Preservation of echinoderms in this way - not previously recorded from the Middle Cambrian - permits detailed analysis of the three-dimensional stereom structure. Identifiable are fragments of stylophorans and eocrinoids. Stem columnals, most likely from eocrinoids, show a wide and advanced morphological variation indicating articulation similar to that of crinoids. The material from the Exsulans Limestone/Kalby marl ( Ptychagnostus gibbus Zone) represents stem-bearing cystoids older than Akadocrinus from Bohemia. The Andrarum Limestone ( Sole-nopleura brachymetopa Zone) contains echinoderm fragments from a higher stratigraphical level, a level correlatable with that from which the oldest North American stem–bearing cystoid, Eustypocystis , has been recorded.     

Ultrascale and microscale growth dynamics of the cidaroid spine of Phyllacanthus imperialis revealed by 26Mg labeling and NanoSIMS isotopic imaging

Growth dynamics of the primary spine of the cidaroid sea urchin Phyllacanthus imperialis was assessed for the first time using pulsed ²⁶Mg‐labeling and NanoSIMS isotopic imaging. The sea urchin was incubated twice (for 48 h) in artificial seawater with elevated level of ²⁶Mg. After each labeling event, the sea urchin was returned for 72 h to seawater with natural isotopic abundance of ²⁶Mg. NanoSIMS ion microprobe was subsequently used to visualize the labeled regions of the spine with submicrometer lateral resolution. The growth of the new skeleton was restricted to the distalmost and peripheral portions of the spine. Skeletogenesis involved mostly the deposition of continuous thickening layers and lateral growth involving bridges between previously formed trabeculae. The timescale of formation of individual thickening layers (ca. 1 µm in width) on the stereom trabeculae was on the order of 1 day. Longitudinal growth occurred mainly at the periphery in the form of small portions of the thickening deposits or more massive microspines that appeared to branch and fuse with those above and below. These microspines were found to grow at about 10 µm/day. These results reveal that the skeletal growth of a juvenile cidaroid spine is complex and highly heterogeneous, with different extension rates depending on the stage of the stereom development and/or direction of the growth fronts. The growth pattern observed here at the submicrometer scale provides direct evidence supporting the earlier suggestions that the lamellar structure of echinoderm stereom is formed by periodic deposition of continuous mineral layers. J. Morphol. 275:788–796, 2014. © 2014 Wiley Periodicals, Inc.     

X-ray microCT study of pyramids of the sea urchin Lytechinus variegatus

This paper reports results of a novel approach, X-ray microCT, for quantifying stereom structures applied to ossicles of the sea urchin Lytechinus variegatus. MicroCT, a high resolution variant of medical CT (computed tomography), allows noninvasive mapping of microstructure in 3-D with spatial resolution approaching that of optical microscopy. An intact pyramid (two demipyramids, tooth epiphyses, and one tooth) was reconstructed with 17 microm isotropic voxels (volume elements); two individual demipyramids and a pair of epiphyses were studied with 9-13 microm isotropic voxels. The cross-sectional maps of a linear attenuation coefficient produced by the reconstruction algorithm showed that the structure of the ossicles was quite heterogeneous on the scale of tens to hundreds of micrometers. Variations in magnesium content and in minor elemental constitutents could not account for the observed heterogeneities. Spatial resolution was insufficient to resolve the individual elements of the stereom, but the observed values of the linear attenuation coefficient (for the 26 keV effective X-ray energy, a maximum of 7.4 cm(-1) and a minimum of approximately 2 cm(-1) away from obvious voids) could be interpreted in terms of fractions of voxels occupied by mineral (high magnesium calcite). The average volume fraction of mineral determined for a transverse slice of the demipyramid near where it joins an epiphysis was 0.46; for a slice 3.3 mm adoral it was 0.70. Local volume fractions of mineral approached 1, and, away from resolvable voids, considerable portions of the demipyramids had volume fractions of calcite at or below approximately 0.33. MicroCT imaging of a demipyramid before and after infiltration with a high absorptivity fluid (sodium polytungstate) confirmed the determination of the volume fractions of minerals.     

The ichnogenus Tremichnus in the Lower Silurian of western New York

Discovery of Tremichnus cysticus Brett on a crinoid pluricolumnal from the Willowvale Shale of western New York extends the range of this ichnogenus into the Lower Silurian. Pits attributable to Tremichnus apparently represent embedment sites of parasites or commensalistic, filter-feeding organisms of unknown affinities. Embedment of T. cysticus may have been accomplished by localized inhibition of stereom growth within the pits aided by secretion of substances that stimulated accretion of stereom by the host pelmatozoan in areas immediately surrounding the pits.     

The role of rock-phosphate-solubilizing fungi and vesicular–arbusular-mycorrhiza (VAM) in growth of wheat plants fertilized with rock phosphate

A total of 36 fungal species isolated from soil were tested for their ability to solubilize rock phosphate (RP) in agar plates. Most of these fungi were non-rock phosphate solubilizers, but two isolates, Aspergillus niger and Penicillium citrinum, had high activity. Liquid culture experiments revealed that both fungi caused a remarkable drop in pH of culture media and solubilized considerable amounts of phosphate. The effects on wheat of inoculation with vesicular–arbuscular mycorrhizal fungi and rock-phosphate-solubilizing fungi and fertilization with rock phosphate were studied in sterilized pot soils, nonsterilized pot trials and in field plot soils. Rock phosphate fertilization and inoculation with Glomus constrictum and rock-phosphate-solubilizing fungi (A. niger and P. citrinum) significantly increased dry matter yield of wheat plants under all experimental conditions. However, the effect was more evident in non- sterilized pot soils and in the field than in sterilized pots. Rock phosphate had no significant effect on the total phosphorus content of plants grown under pot conditions but it was significantly increased in field plots; the effect of inoculation with fungi (G. constrictum, A. niger and P. citrinum) on plant phosphorus was closely related to this in dry matter production. The greatest positive effect on growth and phosphorus contents of wheat plants was recorded in the treatments that received rock phosphate and were inoculated with a mixed inoculum of the three microorganisms used, followed by dual inoculation treatments of G. constrictum plus either A. niger or P. citrinum.     

The role of rock-phosphate-solubilizing fungi and vesicular–arbusular-mycorrhiza (VAM) in growth of wheat plants fertilized with rock phosphate

A total of 36 fungal species isolated from soil were tested for their ability to solubilize rock phosphate (RP) in agar plates. Most of these fungi were non-rock phosphate solubilizers, but two isolates, Aspergillus niger and Penicillium citrinum, had high activity. Liquid culture experiments revealed that both fungi caused a remarkable drop in pH of culture media and solubilized considerable amounts of phosphate. The effects on wheat of inoculation with vesicular–arbuscular mycorrhizal fungi and rock-phosphate-solubilizing fungi and fertilization with rock phosphate were studied in sterilized pot soils, nonsterilized pot trials and in field plot soils. Rock phosphate fertilization and inoculation with Glomus constrictum and rock-phosphate-solubilizing fungi (A. niger and P. citrinum) significantly increased dry matter yield of wheat plants under all experimental conditions. However, the effect was more evident in non- sterilized pot soils and in the field than in sterilized pots. Rock phosphate had no significant effect on the total phosphorus content of plants grown under pot conditions but it was significantly increased in field plots; the effect of inoculation with fungi (G. constrictum, A. niger and P. citrinum) on plant phosphorus was closely related to this in dry matter production. The greatest positive effect on growth and phosphorus contents of wheat plants was recorded in the treatments that received rock phosphate and were inoculated with a mixed inoculum of the three microorganisms used, followed by dual inoculation treatments of G. constrictum plus either A. niger or P. citrinum.     

Do fecal shields provide physical protection to larvae of the tortoise beetles Plagiometriona flavescens and Stolas chalybea against natural enemies?

During their larval phase, Plagiometriona flavescens (Boheman, 1855) and Stolas chalybea (Germar, 1824) (Chrysomelidae: Cassidinae), carry masses of feces and exuviae on their back called fecal shield, and it is suggested to protect larvae against natural enemies. We investigated if the physical barrier provided by the shield plays any role in the defense of these larvae. We conducted a field experiment checking the mortality of larvae of both species with their natural shields substituted by an artificial shield, with shields removed and with their shields intact. Mortality controls for each of the 3 shield treatments were carried out on host plants protected against natural enemies. On both species we observed that larvae with their shields intact had a significant lower mortality proportion than larvae with artificial shields or without shields. Control larvae on protected plants had low mortality. Our results agree with literature data, showing that fecal shields do not provide a physical protection to larvae but are important in their defense, probably due to the chemicals present in them.     

The attachment of collagenous ligament to stereom in primary spines of the sea-urchin, Eucidaris tribuloides

The similar proximal and distal attachments to the stereom of primary spine ligament in the echinoid Eucidaris tribuloides are described, from thin sections and SEM studies on frozen and fractured spine articulations and ligaments from decalcified material. The orthogonal structure of the general stereom is modified on the attachment zones where bundles of collagen cylinders enter approximately hexagonally arranged channels. Straps of collagen extend in parallel series between adjacent bundles via regularly placed ports and collagen loops rather than non-striated 'tendons' pass over skeletal trabeculae. The regular pattern of collagen straps is most evident on the proximal and distal attachment zones. Mechanical features of the non-adhesive mode of attachment are considered, together with similarities and differences between insertion of muscle cells and mutable collagenous tissue (ligament) in echinoderms.     

Sequential study of synaptonemal complexes in mouse spermatocytes by light and electron microscopy

Synaptonemal complex (SC) studies in male mice from the beginning of meiosis to its completion (7–40 days of age) indicate: (1) the existence of a leptotene stage with fragmented and completely unpaired axial elements; (2) that synapsis begins at several different initiation points of the axial elements, resulting in X-shaped and Y-shaped configurations; (3) that interlocking of axial elements at zygotene is a normal phenomenon; and (4) that zygotene (presynaptic) and diplotene (postsynaptic) configurations are different from each other. This investigation received financial support from the special Programme of Research, Development and Research Training in Human Reproduction, World Health Organization.     

Morphometric, AFLP and plastid microsatellite variation in populations of Scalesia divisa and S. incisa (Asteraceae) from the Galápagos Islands

Analysis of the stem vasculature of the American climbing palm Desmoncus reveals structural features differing significantly from the Old World rattan genus Calamus . Desmoncus has a more directly continuous vascular system but nevertheless shows a vessel distribution that makes for high hydraulic resistance in the axial xylem. Desmoncus is like Calamus in having a single very wide metaxylem vessel in each central axial bundle and is also without direct vascular contact between protoxylem and metaxylem tracheary elements. However, in Desmoncus the stem vascular bundle system resembles that in tree palms (as has been described in the model palm Rhapis excelsa ) in having a continuing axial bundle that branches from each outgoing leaf trace together with a large number of bridge connections between leaf traces and peripheral axial bundles. Resistance to axial water transport is, however, evident in the narrowness of the continuing metaxylem elements in the peripheral stem vascular region. Desmoncus has scalariform perforation plates with few thickening bars in the metaxylem vessels, unlike the simple perforation plates found in Calamus . Thus, Desmoncus shows only limited convergence in stem vascular architecture toward the extreme modifications found in Calamus . This is not unexpected since it is clear that the climbing habit evolved independently in the two genera.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society, 2003, 142 , 243−254.     

Axial variation in the three-spine stickleback: genetic and environmental factors

Subtle differences in the pattern of arrangement of types of vertebrae and associated median skeletal structures between a benthic and limnetic species pair of threespine stickleback from Paxton Lake, British Columbia, are typical of those found throughout the range of the Gasterosteus aculeatus species complex. We established laboratory colonies from just three individuals of each species, and studied the effect of three generations of inbreeding on axial morphology. There was sufficient divergence in the location of individual elements between families to regenerate close to the entire range of axial diversity seen in threespine sticklebacks worldwide. Analysis of the patterns of variance and covariance between the axial locations of elements provides evidence for the action of both meristic and homeotic processes in the generation of morphological divergence within each species. Hybrid sticklebacks produced by the cross of limnetic and benthic parents tend to have intermediate morphologies, with dominance of either parental type evident for some elements. Effects of temperature and salinity were found to be small in direct comparison with between-family effects, and varied according to genetic background. These results demonstrate that considerable genetic variation for axial morphology is maintained in natural populations of threespine stickleback, and that differences between populations may be brought about rapidly by changes in frequency of alleles that have coordinated effects along the body axis.