Quantification of porosity in Acropora pulchra (Brook 1891) using X-ray micro-computed tomography techniques

Skeletal density and porosity characteristics are key parameters for investigations into scleractinian coral growth and for assessing the effects of a range of anthropogenic influences on coral reefs. Typically, skeletal density is measured by using planar X-rays of thin slabs cut from mound-shaped colonies or, for branching forms, by using methods based on Archimedean principles. This paper describes a novel non-destructive technique based on micro-computed tomography (micro-CT) to measure porosity of branching coral skeleton. This approach incorporates methods for segmenting internal and external portions of branch and for distinguishing between skeleton and air, whilst accounting for the effects of beam hardening. Measurements were obtained from colonies of branching Acropora pulchra collected across a reef-flat transect at King Reef, central nearshore Great Barrier Reef. The results show significant variation in porosity within and among branches sampled from individual colonies, but not within a reef-flat transect. Micro-CT techniques yield comparable results to traditional methods based on Archimedean principles, but offer advantages in their suitability for a wider range of coral specimens because of the non-destructive nature of the technique and in their more rigorous control of model parameters that can bias results.     

Pincer-like claws in centipedes (Chilopoda): multiple evolutionary origin of similar form and serial pattern

In most Chilopoda, the walking legs end in a single-tip claw usually accompanied by short accessory spines. Instead, in all species of three small and only distantly related geophilomorph taxa (Diphyonyx, Neogeophilidae, Eucratonyx), the claws of an anterior set of leg pairs are unusually pincer-like. By integrating different microscopic techniques, including confocal laser scanning microscopy, we found that these modified claws are very similar in form, internal structure, and pattern of variation in shape along the trunk in all three taxa: the claws are distinctly swollen and bent, provided with peculiar bulges, and flanked by a conspicuous additional branch, either cylindrical or flattened, which overreaches the tip of the claw; instead, the internal cuticular features are not modified with respect to the condition in the other centipedes, claiming against the possibility of controlled abduction/adduction between claw and branch. Irrespective of the total number of leg pairs (63–129), the claws change gradually from pincer-like to usual shape invariantly in the range spanning between the 34 and the 45% of the total number of leg pairs. Despite these similarities, pincer-like claws originated independently in the three taxa, and by way of fundamentally different changes, either by the dramatic modification of the already existent anterior accessory spine (Diphyonyx, Neogeophilidae) or by the production of a novel cuticular projection (Eucratonyx). Moreover, their shared pattern of variation along the body was most probably constrained by already operating developmental processes controlling the longitudinal patterning of the trunk.     

Critical Evaluation of Branch Polarity and Apical Dominance as Dictators of Colony Astogeny in a Branching Coral

The high morphological resemblance between branching corals and trees, can lead to comparative studies on pattern formation traits, best exemplified in plants and in some cnidarians. Here, 81 branches of similar size of the hermatypic coral Stylophora pistillata were lopped of three different genets, their skeletons marked with alizarin red-S, and divided haphazardly into three morphometric treatment groups: (I) upright position; (II) horizontal position, intact tip; and (III) horizontal position, cut tip. After 1 y of in-situ growth, the 45 surviving ramets were brought to the laboratory, their tissues removed and their architectures analyzed by 22 morphological parameters (MPs). We found that within 1 y, isolated branches developed into small coral colonies by growing new branches from all branch termini, in all directions. No architectural dissimilarity was assigned among the three studied genets of treatment I colonies. However, a major architectural disparity between treatment I colonies and colonies of treatments II and III was documented as the development of mirror structures from both sides of treatments II and III settings as compared to tip-borne architectures in treatment I colonies. We did not observe apical dominance since fragments grew equally from all branch sides without documented dominant polarity along branch axis. In treatment II colonies, no MP for new branches originating either from tips or from branch bases differed significantly. In treatment III colonies, growth from the cut tip areas was significantly lower compared to the base, again, suggesting lack of apical dominance in this species. Changes in branch polarity revealed genet associated plasticity, which in one of the studied genets, led to enhanced growth. Different genets exhibited canalization flexibility of growth patterns towards either lateral growth, or branch axis extension (skeletal weight and not porosity was measured). This study revealed that colony astogeny in S. pistillata is a regulated process expressed through programmed events and not directly related to simple energy trade-off principles or to environmental conditions, and that branch polarity and apical dominance do not dictate colony astogeny. Therefore, plasticity and astogenic disparities encompass a diversity of genetic (fixed and flexible) induced responses.     

Correlated evolution of the cis-acting regulatory elements and developmental expression of the Drosophila Gld gene in seven species from the subgroup melanogaster

The tissue-specific expression patterns of glucose dehydrogenase (GLD) exhibit a high degree of inter specific variation in the adult reproductive tract among the species in the genus Drosophila. We chose to focus on the evolution of GLD expression and the evolution of the Gld promoter in seven closely related species in the mela-nogaster subgroup as a means of elucidating the relationship of changes in cis-acting regulatory elements in the Gld promoter region with changes in tissue-specific expression. Although little variation in tissue-specific patterns of GLD was found in nonreproductive tissues during development, a surprisingly high level of variation was observed in the expression of GLD in both developing and ma-ture reproductive organs. In some cases this variation is correlated with changes in sequence elements in the Gld promoter which were previously shown to direct tissue-specific expression in the reproductive tract. In particular D. teissieri adult males do not express GLD in their ejaculatory ducts, atypical of the melanogaster subgroup species. The Gld promoter region of D. teissieri specifically lacks all three of the TTAGA regulatory elements present in D. melanogaster. The TTAGA elements were previously shown to direct reporter gene expression to the ejaculatory duct. Together these data suggest the absence or presence of the TTAGA elements may be responsible for variation in the absence or presence of GLD in the ejaculatory duct among species. © 1994 Wiley-Liss, Inc.     

The Identity of Specimens of the Anastrepha fraterculus Complex (Diptera, Tephritidae) with Atypical Aculeus Tip

Several specimens collected in Paraguay along with Anastrepha fraterculus (sensu lato) have an aculeus tip similar to species from the fraterculus complex, but the teeth of the aculeus of these specimens are poorly defined. As Anastrepha species identification is based mostly on subtle differences in the aculeus tip, we studied these specimens with atypical aculeus tips (with poorly defined teeth) that slightly differs from the aculeus tip of species of the fraterculus complex (with well-developed blunt teeth), to determine if this is due to intraspecific variation or if it can characterize a full species. The Paraguayan specimens were separated in six groups under stereomicroscope according to variation in their aculeus tip. Specimens within each group were studied by means of morphometrics (traditional and geometric) and gene sequence analysis (COI and ITS1). Morphometric analyses were significant, but no clear groups were formed by the discriminant analyses of the aculeus and wing, and the COI and ITS1 sequence analysis clustered specimens with all six aculeus variations. Therefore, the subtle morphological differences observed in the aculeus tip of Paraguayan specimens are intraspecific variations and the Paraguayan specimens were more genetically closely related to Anastrepha sp. 3 from the fraterculus complex.     

Understanding variation in salamander ionomes: A nutrient balance approach

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On the causes of density banding in skeletons of corals of the genus Montastraea

To obtain a reliable climate reconstruction from coral skeletons it is first necessary to understand the way these grow and incorporate information. Thickness of skeletal elements (exothecal and endothecal dissepiments, costae, septa, and theca-wall) as well as the spacing between exothecal and endothecal dissepiments of the four extant Atlantic species of Montastraea (M. annularis, M. faveolata, M. franksi and M. cavernosa) were measured through high and low density bands. Our results show that growth periodicity, controlled by the effect of temperature, is expressed in changes in thickness of costae and exothecal dissepiments in the four studied Montastraea species, with no changes in endothecal elements and theca-wall thickness which, in turn, has implications for research on inclusive records using these species. Spacing between both exothecal and endothecal dissepiments resulted without changes along the high and low density bands, and we found evidence that there is a rhythmical formation of these structures linked somehow to lunar cycles.     

Skeletal development in <Emphasis Type="Italic">Acropora palmata</Emphasis> (Lamarck 1816): a scanning electron microscope (SEM) comparison demonstrating similar mechanisms of skeletal extension in axial versus encrusting growth

Many Acropora palmata colonies consist of an encrusting basal portion and erect branches. Linear growth of the skeleton results in extension along the substrate (encrusting growth), lengthening of branches (axial growth) and thickening of branches and crust (radial growth). Scanning Electron Microscopy is used to compare the mechanisms of skeletal extension between encrusting growth and axial growth. In encrusting growth, the distal margin of the skeleton lacks corallites (which develop about 1 mm from the edge); in contrast, in axial growth, axial corallites along the branch tip form the distal portion of the skeleton. In both locations, the distal margin of the skeleton consists of a lattice-like structure composed of rods that extend from the body of the skeleton and bars that connect these rods. An actively extending skeleton is characterized by sharply pointed rods and partially developed bars. Distal growth of rods (and formation of bars) is effected by the formation of new sclerodermites. Each sclerodermite begins with the deposition of fusiform crystals (that range in length from 1 to 5 μm). These provide a surface for nucleation and growth of spherulitic tufts, clusters of short (<1 μm long) aragonite needles. The needles that are oriented perpendicular to the axis of the skeletal element (rod or bar), and perpendicular to the overlying calicoblastic epithelium, continue extension to appear on the surface of the skeleton as 10–15 μm wide bundles (of needle tips) called fasciculi. However, some crusts that abut competitors for space have a different morphology of skeletal elements (rods and bars). The distal edge of these crusts terminates in blunt coalescing rods, and bars that are fully formed. Absence of fusiform crystals, lack of sharply pointed rods and bars, and full development of sclerodermites characterize a skeletal region that has ceased, perhaps only temporarily, skeletal extension.     

Skeletal variation related to arm regeneration in Metacrinus and Saracrinus, Recent stalked crinoids

Variations in the number of skeletal elements in the proximal portion of the arms (primibrachials) of Metacrinus rotundus Carpenter and Saracrinus nobilis (Carpenter) are compared. Older specimens of Metacrinus rotundus show increased variation in number of elements along with a tendency toward reduction, whereas the number of elements remains nearly constant in Saracrinus nobilis. The variation in Metacrinus rotundus is attributed to inaccurate arm regeneration following autotomy. Separation of Saracrinus from Metacrinus on the basis of number of skeletal elements is therefore possible, but caution must be used in the application of taxonomic characters which might be influenced by regeneration.     

自然干旱梯度下的酸枣表型变异

表型变异是植物应对环境变化的一种策略。酸枣植物从中国东部沿海到内陆腹地均有分布,其表型性状的变异可能解释其对自然干旱梯度的适应机制。为验证这一假说,以烟台、石家庄、银川、吐鲁番4个自然干旱梯度生境中生长的酸枣三年生植株的12个表型性状为调查研究对象,采用变异系数和巢式方差分析对酸枣的表型变异进行分析。结果表明:(1)从烟台到银川,叶面积、叶长、叶周长和叶柄长总体呈减小的趋势,而比叶面积呈增大的趋势;(2)随着干旱程度的增强,二次枝的长度、二次枝的基部粗、二次枝的枣吊数、茎比密度和茎水分含量均呈减小趋势,并且种子重和种子短轴长也均呈减小的趋势;(3)对沿干旱梯度分布的4个酸枣种群而言,叶性状的平均变异系数(33.70%)枝性状的平均变异系数(32.41%)种子性状的平均变异系数(9.07%),并且酸枣性状间存在很强的协变。结果表明酸枣的地上部分形态性状沿干旱梯度表现出很强的变异,推测在未来的气候变化下,酸枣将通过这种表型变异的有效组合来适应环境变化。     

USING HITCHHIKING GENES TO STUDY ADAPTATION AND DIVERGENCE DURING SPECIATION WITHIN THE DROSOPHILA MELANOGASTER SPECIES COMPLEX

Several studies of intraspecific and interspecific DNA sequence variation from Drosophila loci have revealed a pattern of low intraspecific variation from genomic regions of low recombination. The mechanisms consistently invoked to explain these patterns are the selective sweep of advantageous mutations together with genetic hitchhiking of linked loci. To examine the effect of selective sweeps on genetic divergence during speciation, we studied two loci in different genomic regions thought to be subject to selective sweeps. We obtained DNA sequences from 1.1kb pair portions of the fourth chromosome locus cubitus interruptus Dominant (ci^D) and from the asense locus near the telomere of the X chromosome. At ci^D, we found very low variation among multiple lines of Drosophila mauritiana and D. sechellia. This finding is consistent with an earlier report of very low variation in D. melanogaster and D. simulans at ci^D and supports the conclusion of selective sweeps and genetic hitchhiking on the nonrecombining fourth chromosome. The pattern of variation found at asense suggests that a selective sweep has occurred recently at the tip of the X chromosome in D. simulans, but not in D. melanogaster or D. mauritiana. The data from ci^D and asense are compared with data from three X chromosome loci (period, zeste, and yolk protein 2) that experience normal levels of recombination. By examining estimated genealogies and the rates at which different classes of mutations have accumulated, we conclude that selective sweeps are common occurrences on the fourth chromosome but less common near the tip of the X chromosome. An interesting pattern of low variation at ci^D among D. simulans, D. mauritiana, and D. sechellia suggests that a selective sweep may have occurred among these forms even after divergence into separate species had begun.     

Nitrogen allocation and carbon isotope fractionation in relation to intercepted radiation and position in a young Pinus radiata D. Don tree

The three dimensional distribution of intercepted radiation, intercellular CO₂ concentration (C_i) and late summer needle nitrogen (N) concentration were determined at the tips of all 54 branches in a 6·2-m-tall Pinus radiata D. Don tree growing in a New Zealand plantation. Measurements included above- and below-canopy irradiance, leaf stable carbon isotopic composition (δ¹³C) and tree canopy architecture. The radiation absorption component of the model, MAESTRO, was tested on site and then used to determine the branch tip distribution of intercepted radiation. We hypothesized that in branch tip needles: (i) the allocation of nitrogen and other nutrients would be closely associated with the distribution of intercepted radiation, reflecting carbon gain optimization theory, and (ii) C_i would predominantly reflect changes in photosynthetic rate (A) rather than stomatal conductance (g_s), indicating that the increase in A for a given increase in N concentration was larger than the corresponding increase in g_s. Needle nitrogen concentration was poorly related to intercepted radiation, regardless of the period over which the latter was calculated. At a given height, there was a large azimuthal variation in intercepted radiation but N concentration was remarkably uniform around the tree canopy. There was, however, a linear and positive correspondence between N concentration and δ¹³C and needle height above ground (r² = 0·73 and 0·68, respectively). The very strong linear correspondence between N concentration and C_i (r² = 0·71) was interpreted, using gas exchange measurements, as supporting our second hypothesis. Recognizing the strong apical control in P. radiata and possible effects of leaf nitrogen storage in an evergreen species, we propose that the tree leader must have constituted a very strong carbon sink throughout the growing season, and that the proximity of branch tip needles to the leader affected their photosynthetic capacity and nutrient concentration, independent of intercepted radiation. This implies an integrated internal determination of resource allocation within the tree and challenges the current convention that resources are optimally distributed according to the profile of intercepted radiation.     

Dynamics of presynaptic protein recruitment induced by local presentation of artificial adhesive contacts

In this study, we introduce a novel approach to induce and observe the formation of presynaptic compartments in axons through a combination of atomic force microscopy (AFM) and fluorescence microscopy. First, we use a poly‐D ‐lysine‐coated bead attached to an AFM tip to induce the recruitment of two synaptic proteins, bassoon and synaptophysin, and measure their absolute arrival times to the presynaptic department. We find that bassoon arrives before synaptophysin. Second, we observe the formation of very long (several 10s of μm), structured, protein‐containing membranous strings as the AFM tip was withdrawn from the axon. It is conceivable that these strings might be a novel mechanism by which new neurites or branch points along existing neurites may be generated in situ. © 2012 Wiley Periodicals, Inc. Develop Neurobiol, 2013     

Functional anatomy and adaptation of the third to sixth thoracic vertebrae in primates using three-dimensional geometric morphometrics

The morphology of the cranial thoracic vertebrae has long been neglected in the study of primate skeletal functional morphology. This study explored the characteristics of the third to sixth thoracic vertebrae among various positional behavioural primates. A total of 67 skeletal samples from four species of hominoids, four of cercopithecoids, and two of platyrrhines were used. Computed tomography images of the thoracic vertebrae were converted to a three-dimensional (3D) bone surface, and 104 landmarks were obtained on the 3D surface. For size-independent shape analysis, the vertebrae were scaled to the same centroid size, and the normalised landmarks were registered using the generalised Procrustes method. Principle components of shape variation among samples were clarified using the variance–covariance matrix of the Procrustes residuals. The present study revealed that the transverse processes were more dorsally positioned in hominoids compared to non-hominoids. The results showed that not only a dorsolaterally oriented but also a dorsally positioned transverse process in relation to the vertebral arch contribute to the greater dorsal depth in hominoids than in monkeys. The thoracic vertebrae of Ateles and Nasalis show relatively dorsoventrally low and craniocaudally long vertebrae with craniocaudally long zygapophyses and craniocaudally long base/short tip of the caudally oriented spinous process, accompanied by a laterally oriented and craniocaudally long base of the transverse process. Despite being phylogenetically separated, the vertebral features of Ateles (suspensory platyrrhine with its prehensile tail's aid) are similar to those of Nasalis (arboreal quadrupedal/jumping/arm-swing colobine). The morphology of the third to sixth thoracic vertebrae tends to reflect the functional adaptation in relation to positional behaviour rather than the phylogenetic characteristics of hominoids, cercopithecoids, and platyrrhines.

     

Diurnal patterns of skeleton formation in Pocillopora damicornis (Linnaeus)

Scanning, transmission and X-ray microanalytical electron microscopy were used to investigate the skeleton, organic matrix and calicoblastic ectoderm of the reef coral Pocillopora damicornis over a diurnal cycle. All skeletal surfaces, both during day and hight, are fasciculate except for skeletal spines on the branch tip apex at night where small (0.5 m) fusiform crystals are deposited. X-ray microanalysis shows that the fusiform crystals and needle-shaped crystals that compose the fasciculi are distinct forms of calcium carbonate. Demineralization of the skeleton reveals an organic matrix with two components which are related to the formation of fusiform crystals and fasciculi. During the day the calicoblastic ectoderm overlying all skeletal surfaces is 1–3 m thick. At night ultrastructural evidence suggests that skeletal deposition occurs only on those skeletal spines at the branch tip apex which are growing parallel with the branch growth axis. The calicoblastic ectoderm overlying apical skeletal spines at night shows a greater degree of cellular activity, and is thicker, than calicoblastic ectoderm overlying both other skeletal surfaces at night (<8 m cf. >6 m) and all skeletal surfaces during the day (<8 m cf. >3 m). The deposition of fusiform crystals on skeletal spines at the branch tip apex is proposed to promote deposition of fasciculi during the day, relative to other skeletal surfaces, providing a mechanism determining apical growth of branch tips. The results are discussed with respect to previous concepts of skeletal deposition in scleractinian corals.     

A highly polymorphic region in the X chromosome of Drosophila

Summary Many cloned regions of the Drosophila genome show minimal variation between strains in overall sequence arrangement. While restriction site polymorphisms occur and the location of transposable elements may vary from one strain to another, such changes appear to be relatively minor variations, superimposed on overall genome stability. In contrast to this general situation, we describe here a segment of the X chromosome that is highly polymorphic in four strains of D. melanogaster and in D. simulans. The strains differ in the presence and extent of a short duplication and the presence of repetitive DNA. These results suggest that different regions of the genome may be subject to different evolutionary constraints, with some regions being particularly prone to extensive changes, even within a single species.     

Appendicular skeletal morphology in minute salamanders,genus Thorius (amphibia: Plethodontidae): Growth regulation,adult size determination,and natural variation

Patterns of growth and variation of the appendicular skeleton were examined in Thorius, a speciose genus of minute terrestrial plethodontid salamanders from southern Mexico. Observations were based primarily on ontogenetic series of each of five species that collectively span the range of adult body size in the genus; samples of adults of each of seven additional species provided supplemental estimates of the full range of variation of limb skeletal morphology. Limbs are generally reduced, i.e., pedomorphic, in both overall size and development, and they are characterized by a pattern of extreme variation in the composition of the limb skeleton, especially mesopodial elements, both within and between species. Fifteen different combinations of fused carpal or tarsal elements are variably present in the genus, producing at least 18 different overall carpal or tarsal arrangements, many of which occur in no other plethodontid genus. As many as four carpal or tarsal arrangements were observed in single population samples of each of several; five tarsal arrangements were observed in one population of T. minutissimus. Left-right asymmetry of mesopodial arrangement in a given specimen is also common. In contrast, several unique, nonpedomorphic features of the limb skeleton, including ossification of the typically cartilaginous adult mesopodial elements and ontogenetic increase in the degree of ossification of long bones, are characteristic of all species and distinguish Thorius from most related genera. They form part of a mechanism of determinate skeletal growth that restricts skeletal growth after sexual maturity. Interspecific differences in the timing of the processes of appendicular skeletal maturation relative to body size are well correlated with interspecific differences in mean adult size and size at sexual maturity, suggesting that shifts in the timing of skeletal maturation provide a mechanism of achieving adult size differentiation among species. Processes of skeletal maturation that confer determinate skeletal growth in Thorius are analogous to those typical of most amniotes – both groups exhibit ontogenetic reduction and eventual disappearance of the complex of stratified layers of proliferating and maturing cartilage in long bone epiphyses – but, unlike most amniotes, Thorius lacks secondary ossification centers. Thus, the presence of secondary ossification centers cannot be used as a criterion for establishing determinate skeletal growth in all vertebrates.