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At least 109 records · Page 6

An amphioxus nodal gene (AmphiNodal) with early symmetrical expression in the organizer and mesoderm and later asymmetrical expression associated with left-right axis formation

The full-length sequence and zygotic expression of an amphioxus nodal gene are described. Expression is first detected in the early gastrula just within the dorsal lip of the blastopore in a region of hypoblast that is probably comparable with the vertebrate Spemann's organizer. In the late gastrula and early neurula, expression remains bilaterally symmetrical, limited to paraxial mesoderm and immediately overlying regions of the neural plate. Later in the neurula stage, all neural expression disappears, and mesodermal expression disappears from the right side. All along the left side of the neurula, mesodermal expression spreads into the left side of the gut endoderm. Soon thereafter, all expression is down-regulated except near the anterior and posterior ends of the animal, where transcripts are still found in the mesoderm and endoderm on the left side. At this time, expression also begins in the ectoderm on the left side of the head, in the region where the mouth later forms. These results suggest that amphioxus and vertebrate nodal genes play evolutionarily conserved roles in establishing Spemann's organizer, patterning the mesoderm rostrocaudally and setting up the asymmetrical left-right axis of the body.

Non-NASA Center↗

The mammalian retina as a clock

Many physiological, cellular, and biochemical parameters in the retina of vertebrates show daily rhythms that, in many cases, also persist under constant conditions. This demonstrates that they are driven by a circadian pacemaker. The presence of an autonomous circadian clock in the retina of vertebrates was first demonstrated in Xenopus laevis and then, several years later, in mammals. In X. laevis and in chicken, the retinal circadian pacemaker has been localized in the photoreceptor layer, whereas in mammals, such information is not yet available. Recent advances in molecular techniques have led to the identification of a group of genes that are believed to constitute the molecular core of the circadian clock. These genes are expressed in the retina, although with a slightly different 24-h profile from that observed in the central circadian pacemaker. This result suggests that some difference (at the molecular level) may exist between the retinal clock and the clock located in the suprachiasmatic nuclei of hypothalamus. The present review will focus on the current knowledge of the retinal rhythmicity and the mechanisms responsible for its control.

Review Literature↗

Retinoid quantification by HPLC/MS(n)

Retinoic acid (RA) mediates most of the biological effects of vitamin A that are essential for vertebrate survival. It acts through binding to receptors that belong to the nuclear receptor transcription factor superfamily (Mangelsdorf et al. 1994). It is also a highly potent vertebrate teratogen. To determine the function and effects of endogenous and exogenous RA, it is important to have a highly specific, sensitive, accurate, and precise analytical procedure. Current analyses of RA and other retinoids are labor intensive, of poor sensitivity, have limited specificity, or require compatibility with RA reporter cell lines (Chen et al. 1995. BIOCHEM: Pharmacol. 50: 1257-1264; Creech Kraft et al. 1994. BIOCHEM: J. 301: 111-119; Lanvers et al. 1996. J. Chromatogr. B Biomed. Appl. 685: 233-240; Maden et al. 1998. DEVELOPMENT: 125: 4133-4144; Wagner et al. 1992. DEVELOPMENT: 116: 55-66). This paper describes an HPLC/mass spectrometry/mass spectrometry product ion scan (HPLC/MS(n)) procedure for the analysis of retinoids that employs atmospheric pressure chemical ionization MS. The retinoids are separated by normal-phase column chromatography with a linear hexane-isopropanol-dioxane gradient. Each retinoid is detected by a unique series of MS(n) functions set at optimal collision-induced dissociation energy (30% to 32%) for all MS(n) steps. The scan events are divided into three segments, based on HPLC elution order, to maximize the mass spectrometer duty cycle. The all-trans, 9-cis, and 13-cis RA isomers are separated, if desired, by an isocratic hexane-dioxane-isopropanol mobile phase. This paper describes an HPLC/MS(n) procedure possessing high sensitivity and specificity for retinoids.

NASA Discipline Developmental Biology↗

Cardiovascular responses of snakes to hypergravity

Snakes have provided useful vertebrate models for understanding circulatory adaptation to gravity, attributable to their elongate body shape and evolutionary diversificaton in terms of ecology and behavior. Recently we have studied cardiovascular responses of snakes to hypergravic acceleration forces produced acutely in the head-to-tail direction (+Gz) on a short-arm centrifuge. Snakes were held in a nearly straight position within a horizontal plastic tube and subjected to a linear force gradient during acceleration. Carotid blood flow provided an integrated measure of cardiovascular performance. Thus, cardiovascular tolerance of snakes to stepwise increments of Gz was measured as the caudal Gz force at which carotid blood flow ceased. Tolerance to increasing Gz varies according to adaptive evolutionary history inferred from the ecology and behavior of species. With respect to data for six species we investigated, multiple regression analysis demonstrates that Gz tolerance correlates with gravitational habitat, independently of body length. Relative to aquatic and non-climbing species, carotid blood flow is better maintained in arboreal or scansorial species, which tolerate hypergravic forces of +2 to +3.5 Gz. Additionally, semi-arboreal rat snakes (Elaphe obsoleta) exhibit plasticity of responses to long-term, intermittent +1.5 Gz stress. Compared to non-acclimated controls, acclimated snakes show greater increases of heart rate during head-up tilt or acceleration, greater sensitivity of arterial pressure to circulating catecholamines, higher blood levels of prostaglandin ratios favorable to maintenance of arterial blood pressure, and medial hypertrophy in major arteries and veins. As in other vertebrates, Gz tolerance of snakes is enhanced by acclimation, high arterial pressure, comparatively large blood volume, and body movements. Vascular studies of snakes suggest the importance to acclimation of local responses involving vascular tissue, in addition to centrally mediated responses to fluid shifts.

Review, Tutorial↗

Evolutionary dynamics of plants and animals: a comparative approach

Patterns of longevity and rate of appearance of taxa in the fossil record indicate a different evolutionary dynamic between land plants and marine invertebrates. Among marine invertebrates, rates of taxonomic turnover declined through the Phanerozoic, with increasingly extinction-resistant, long-lived, clades coming to dominate. Among terrestrial vascular plants, rates of turnover increased through the Phanerozoic, with short-lived, extinction-prone clades coming to dominate from the Devonian to the present. Terrestrial vertebrates appear to approximate the marine invertebrate pattern more closely than the plant record. We identify two features which individually or jointly may have influenced this distinction. First, land plants continuously invaded stressful environments during their evolution, while marine invertebrates and terrestrial vertebrates did not. Second, the relative structural simplicity and indeterminate mode of plant growth vs. the relative structural complexity and determinate mode of animal growth may have influenced the timing of major clade origin in the two groups.

Review↗

Otoconia biogenesis, phylogeny, composition and functional attributes

This work consolidates data about these interesting organic crystals of vertebrate inner ears. It addresses 5 aspects of inner ear otoliths not completely understood to date: 1) embryological data that explains the formation of the crystals, 2) the significance of the organic and the inorganic phase of the otolith and the changing patterns of otoconia formation along the evolutionary tree, 3) otoliths contribution for detecting linear acceleration, 4) the effect that altered gravity and aminoglycosides have on the development and adult shape of the crystals, and the evolutionary significance of a changing shape of the crystals from primitive forms (lamprey) to high vertebrate birds and mammals is discussed, 5) functional attributes of the otolithic organs and morphological modifications of the otoliths by physical and chemical insults are presented with an extensive discussion of the most relevant literature published and available to us.

Non-NASA Center↗

Mineral and organic matrix interaction in normally calcifying tendon visualized in three dimensions by high-voltage electron microscopic tomography and graphic image reconstruction

To define the ultrastructural accommodation of mineral crystals by collagen fibrils and other organic matrix components during vertebrate calcification, electron microscopic 3-D reconstructions were generated from the normally mineralizing leg tendons from the domestic turkey, Meleagris gallopavo. Embedded specimens containing initial collagen mineralizing sites were cut into 0.5-micron-thick sections and viewed and photographed at 1.0 MV in the Albany AEI-EM7 high-voltage electron microscope. Tomographic 3-D reconstructions were computed from a 2 degree tilt series of micrographs taken over a minimum angular range of +/- 60 degrees. Reconstructions of longitudinal tendon profiles confirm the presence of irregularly shaped mineral platelets, whose crystallographic c-axes are oriented generally parallel to one another and directed along the collagen long axes. The reconstructions also corroborate observations of a variable crystal length (up to 170 nm measured along crystallographic c-axes), the presence of crystals initially in either the hole or overlap zones of collagen, and crystal growth in the c-axis direction beyond these zones into adjacent overlap and other hole regions. Tomography shows for the first time that crystal width varies (30-45 nm) but crystal thickness is uniform (approximately 4-6 nm at the resolution limit of tomography); more crystals are located in the collagen hole zones than in the overlap regions at the earliest stages of tendon mineralization; the crystallographic c-axes of the platelets lie within +/- 15-20 degrees of one another rather than being perfectly parallel; adjacent platelets are spatially separated by a minimum of 4.2 +/- 1.0 nm; crystals apparently fuse in coplanar alignment to form larger platelets; development of crystals in width occurs to dimensions beyond single collagen hole zones; and a thin envelope of organic origin may be present along or just beneath the surfaces of individual mineral platelets. Implicit in the results is that the formation of crystals occurs at different sites and times by independent nucleation events in local regions of collagen. These data provide the first direct visual evidence from 3-D imaging describing the size, shape, orientation, and growth of mineral crystals in association with collagen of a normally mineralizing vertebrate tissue. They support concepts that c-axial crystal growth is unhindered by collage hole zone dimensions, that crystals are organized in the tendon in a series of generally parallel platelets, and that crystal growth in width across collagen fibrils may follow channels or grooves formed by adjacent hole zones in register.

Non-NASA Center↗

Molecular Mechanisms of Circadian Regulation During Spaceflight

The physiology of both vertebrates and invertebrates follows internal rhythms coordinated in phase with the 24-hour daily light cycle. This circadian clock is governed by a central pacemaker, the suprachiasmatic nucleus (SCN) in the brain. However, peripheral circadian clocks or oscillators have been identified in most tissues. How the central and peripheral oscillators are synchronized is still being elucidated. Light is the main environmental cue that entrains the circadian clock. Under the absence of a light stimulus, the clock continues its oscillation in a free-running condition. In general, three functional compartments of the circadian clock are defined. The vertebrate retina contains endogenous clocks that control many aspects of retinal physiology, including retinal sensitivity to light, neurohormone synthesis (melatonin and dopamine), rod disk shedding, signalling pathways and gene expression. Neurons with putative local circadian rhythm generation are found among all the major neuron populations in the mammalian retina. In the mouse, clock genes and function are more localized to the inner retinal and ganglion cell layers. The photoreceptor, however, secrete melatonin which may still serve a an important circadian signal. The reception and transmission of the non-visual photic stimulus resides in a small subpopulation (1-3%) or retinal ganglion cells (RGC) that express the pigment melanopsin (Opn4) and are called intrisically photoreceptive RGC (ipRGC). Melanopsin peak absorption is at 420 nm and all the axons of the ipRGC reach the SCN. A common countermeasure for circadian re-entrainment utilizes blue-green light to entrain the circadian clock and mitigate the risk of fatigue and health and performance decrement due to circadian rhythm disruption. However, an effective countermeasure targeting the photoreceptor system requires that the basic circadian molecular machinery remains intact during spaceflight. We hypothesize that spaceflight may affect ipRGC and melanopsin expression, which may be a contributing cause of circadian disruption during spaceflight.

Zanello, S. B.↗

Effects of Long Duration Spaceflight on Venous and Arterial Compliance

The visual impairment and intracranial pressure syndrome (VIIP) is a newly described space flight-associated medical condition made up of a constellation of symptoms affecting at least 34% of American astronauts who have flown International Space Station (ISS) missions. VIIP is defined primarily by visual acuity deficits and anatomical changes to eye structures, and is thought to be related to elevated intracranial pressure secondary to space flightinduced cephalad fluid shifts. Loss of visual acuity could be a significant threat to crew health and performance and may be suggestive of other adaptations with implications for years post-flight. Our primary objective is to determine whether vascular compliance is altered by space flight and whether such adaptations are related to the incidence of VIIP. In particular, we will measure ocular parameters and vascular compliance in vessels of the head and neck in astronauts who have no space flight experience, in astronauts before, during, and after space flight, and in bed rest subjects with conditions similar to space flight. Additionally, we will analyze astronaut data from the Lifetime Surveillance of Astronaut Health (LSAH) archive to determine which factors might be predictive of the development of VIIP. The project will be conducted in four separate but related parts. To understand the baseline condition of astronauts without any prior space flight experience, we will study 10 astronauts who have never flown in space by performing a comprehensive evaluation of the vasculature of the head, neck and eyes. Hemodynamic data (stroke volume and blood pressure), ocular (tonometry and ocular ultrasound), venous and arterial parameters will be acquired across a range of tilt angles (20, 10, 0, -10, -20 degrees). Vessels to be studied include the temporal, jugular, and vertebral veins and the cerebral, carotid and vertebral arteries. Ophthalmic data from the annual physical will be obtained through data sharing. To examine the relation between vascular compliance in the head and neck and the development of VIIP after a long duration space flight, we will study 10 astronauts before, during, and after long-duration ISS missions. Pre- and post-flight testing will be identical to that described above. During flight, images of the same vessels of interest will be obtained for later analysis. Ophthalmic data including VIIP scores will be obtained through data sharing from medically-required tests. To investigate the effects of age and elevated sodium intake, two potential contributors to VIIP, we will study 24 men (in two age groups: 25-35 and 45-55) during a 14 day 6deg head-down bed rest, a well-accepted analog of space flight. Standard NASA bed rest conditions will be maintained except for dietary sodium. Sodium intake will be similar to that of ISS astronauts, which is higher than consumed in previous bed rest studies. Pre- and post-bed rest testing procedures will be identical to the testing protocol described above for astronauts. Ophthalmic testing (optical coherence tomography, fundoscopy, and tonometry) will be conducted on the same day that vascular compliance measures are obtained. To identify parameters that may relate to an increase in an astronaut's susceptibility to developing VIIP, we will use data mining techniques to evaluate astronaut data obtained from the LSAH. Medical history, family history, space flight history and its related exposures, and history of high performance jet aircraft exposure will be examined for their potential relationship to ocular data. We hypothesize that the cephalad fluid shift induced by space flight will result in structural and functional adaptations in head and neck vessels leading to decreased vascular compliance and related to the development of VIIP symptoms. Further, although VIIP has not been observed in previous bed rest studies, we hypothesize that an elevated sodium intake will increase the incidence of VIIP symptoms in this space flight analog. Finally, we hypothesize that data mining analyses will reveal relationships between health history, previous exposures (including space flight and high performance aircraft), and the development of VIIP in the astronaut population.

Ribeiro, L. C.↗

NASAs VESGEN: Systems Analysis of Vascular Phenotypes from Stress and Other Signaling Pathways Using GeneLab.

One fundamental requirement shared by humans with all higher terrestrial life forms, including insect wings, higher land plants and other vertebrates, is a complex, fractally branching vascular system. NASA's VESsel GENeration Analysis (VESGEN) software maps and quantifies vascular trees, networks, and tree-network composites according to weighted physiological rules such as vessel connectivity, tapering and bifurcational branching. According to fluid dynamics, successful vascular transport requires a complex distributed system of highly regulated laminar flow. Microvascular branching rules within vertebrates, dicot leaves and the other organisms therefore display many similarities. One unifying perspective is that vascular patterning offers a useful readout that necessarily integrates complex molecular signaling pathways. VESGEN has elucidated changes in vascular pattern resulting from inflammatory, stress response, developmental and other signaling within numerous tissues and major model organisms studied for Space Biology. For a new VESGEN systems approach, we analyzed differential gene expression in leaves of Arabidopsis thaliana reported by GeneLab (GLDS-7) for spaceflight. Vascular-related changes in leaf gene expression were identified that can potentially be phenocopied by mutants in ground-based experiments. To link transcriptional, protein and other molecular change with phenotype, alterations in the Euclidean and dynamic dimensions (x,y,t) of vascular patterns for Arabidopsis leaves and other model species are being co-localized with signaling patterns of single molecular expression analyzed as information dimensions (i,j,k,...). Previously, Drosophila microarray data returned from space suggested significant changes in genes related to wing venation development that include EGF, Notch, Hedghog, Wingless and Dpp signaling. Phenotypes of increasingly abnormal ectopic wing venation in the (non-spaceflight) Drosophila wing generated by overexpression of a Notch antagonist were analyzed by VESGEN. Other VESGEN research applications include the mouse retina, GI and coronary vessels, avian placental analogs and translational studies in the astronaut retina related to health challenges for long-duration missions.

Venation↗

Using NASA's GeneLab for VESGEN Systems Analysis of Vascular Phenotypes from Stress and Other Signaling Pathways

One fundamental requirement shared by humans with all higher terrestrial life forms, including other vertebrates, insects, and higher land plants, is a complex, fractally branching vascular system. NASA's VESsel GENeration Analysis (VESGEN) software maps and quantifies vascular trees, networks, and tree-network composites according to weighted physiological rules such as vessel connectivity, tapering and bifurcational branching. According to fluid dynamics, successful vascular transport requires a complex distributed system of highly regulated laminar flow. Microvascular branching rules within vertebrates, dicot leaves and the other organisms therefore display many similarities. A unifying perspective is that vascular patterning offers a useful readout of molecular signaling that necessarily integrates these complex pathways. VESGEN has elucidated changes in vascular pattern resulting from inflammatory, developmental and other signaling within numerous tissues and major model organisms studied for Space Biology. For a new VESGEN systems approach, we analyzed differential gene expression in leaves of Arabidopsis thaliana reported by GeneLab (GLDS-7) for spaceflight. Vascularrelated changes in leaf gene expression were identified that can potentially be phenocopied by mutants in ground-based experiments. To link transcriptional, protein and other molecular change with phenotype, alterations in the spatial and dynamic dimensions of vascular patterns for Arabidopsis leaves and other model species are being co-localized with signaling patterns of single molecular expression analyzed as information dimensions. Previously, Drosophila microarray data returned from space suggested significant changes in genes related to wing venation development that include EGF, Notch, Hedghog, Wingless and Dpp signaling. Phenotypes of increasingly abnormal ectopic wing venation in the (non-spaceflight) Drosophila wing generated by overexpression of a Notch antagonist were analyzed by VESGEN. Other VESGEN research applications include the mouse retina, GI and coronary vessels, avian placental analogs and translational studies in the astronaut retina related to health challenges for long-duration missions.

Venation↗

A Diverse Mammal-Dominated, Footprint Assemblage from Wetland Deposits in the Lower Cretaceous of Maryland

A newly discovered assemblage of predominantly small tracks from the Cretaceous Patuxent Formation at NASA’s Goddard Space Flight Center, Maryland, reveals one of the highest track densities and diversities ever reported (~70 tracks, representing at least eight morphotypes from an area of only ~2 m(exp 2)). The assemblage is dominated by small mammal tracks including the new ichnotxon Sederipes goddardensis, indicating sitting postures. Small crow-sized theropod trackways, the first from this unit, indicate social trackmakers and suggest slow-paced foraging behavior. Tracks of pterosaurs, and other small vertebrates suggest activity on an organic-rich substrate. Large well-preserved sauropod and nodosaurs tracks indicate the presence of large dinosaurs. The Patuxent Formation together with the recently reported Angolan assemblage comprise the world’s two largest Mesozoic mammal footprint assemblages. The high density of footprint registration at the NASA site indicates special preservational and taphonomic conditions. These include early, penecontemporaneous deposition of siderite in organic rich, reducing wetland settings where even the flesh of body fossils can be mummified. Thus, the track-rich ironstone substrates of the Patuxent Formation, appear to preserve a unique vertebrate ichnofacies, with associated, exceptionally-preserved body fossil remains for which there are currently no other similar examples preserved in the fossil record.

Ray Stanford↗

Murine Spine Composition and Mechanics are Maintained After 30-Days of Spaceflight

As NASA prepares to send astronauts on deep space missions, understanding the detrimental effects of spaceflight on the body is imperative. Spaceflight is known to negatively affect the spine, with astronauts experiencing systemic bone loss and increased post-flight risk of intervertebral disc herniation. Space investigations with rodent models have characterized some of the effects of spaceflight on the mammalian body, but variable experimental conditions have made it difficult to cohesively interpret results. These conditions include housing, flight duration, and for the spine, anatomical location. Thus, the objective of this study was to comprehensively characterize the effects of spaceflight on the bulk properties of the murine lumbar spine. To do this, we examined the lumbar spine of mice flown on the NASA Rodent Research 10 mission and analyzed disc biochemical content, disc joint material properties, lumbar vertebral body microarchitecture, and proximal tibia microarchitecture. Finally, we qualitatively examined bone growth in the vertebral bodies using confocal microscopy and on-orbit injections of calcein. Contrary to prior results, we found little to no effect of spaceflight on lumbar spine properties. Spaceflight, however, did negatively affect both cortical and trabecular bone properties of the proximal tibia load-bearing site. We hypothesize that this may be due to loading associated with increased physical activity in the second half of the mission, observed on in-flight videos, that reload the lumbar spine as mice utilize quadruped ambulation. This increased activity occurs after an initial period of relative inactivity in microgravity. In support of this hypothesis, we observed new L5 bone formation in the last seven days of spaceflight, suggesting that current 30-day experimental flight conditions may not be as effective at lumbar spine unloading as previous shorter flights.

mouse↗

Regional and Sexual Dimorphism in Murine Skeletal Responses to Osteocytic HIF Pathway Modulation

Hypoxia-inducible factors (HIFs) are transcription factors stabilized under hypoxia and degraded under normoxia by the E3 ubiquitin ligase Von Hippel-Lindau (Vhl). In osteocytes, the central orchestrators of bone homeostasis, Vhl and HIFs promote an osteoanabolic transcriptional program. In this study, we assessed unique impacts of osteocytic Vhl deletion versus individual HIF-α paralog stabilization on bone structure, mineralization, and mechanics as a function of sex and skeletal region. Microcomputed tomography revealed that osteocytic Vhl deletion robustly increased trabecular bone mass in both sexes and at both axial and appendicular regions, while HIF-2α accumulation increased femoral metaphyseal bone mass in both sexes while enhancing vertebral bone mass only in males. Vhl deletion paradoxically reduced mineral heterogeneity in male vertebrae, despite raising peak and mean calcium content in both sexes. In contrast, HIF-2α stabilization impaired cortical mineralization and mechanics, especially in females. While cortical mineralization was disrupted in both genotypes, Vhl deletion improved whole bone mechanical properties, suggesting that enhanced geometry and mass offset compromised tissue quality. HIF-1α stabilization exerted negligible impacts on any outcome.

Biological and medical sciences↗

Evaluation of common trap types for capturing wild pigs

Abstract Wild pigs ( Sus scrofa ) are one of the most ecologically harmful invasive vertebrates globally. Due to their negative impacts, wild pig populations are managed via trapping, yet studies investigating various trap designs have been limited in duration, location, and number of trappers. Using data from professional trappers across the southeastern U.S., we evaluated the performance of 3 wild pig trap types (corral, drop, passive net), and the effect of forage season (barren, green‐up, masting) on trapping success. Specifically, we compared trap effectiveness (estimated proportion of targeted wild pigs caught per capture event), trap night effort (number of days to first catch), and the amount of bait and time used per individual take over the lifetime of each trap location across the 3 trap types and forage seasons. Across >850 capture events by 31 trappers, all 3 trap types evaluated were effective at capturing entire social groups of wild pigs. Few differences were observed among trap types, although drop traps in masting season captured a greater estimated proportion of wild pigs per capture event compared to other trap type‐season combinations. Our data also revealed that passive net traps required slightly more bait/wild pig captured than drop traps but there was no difference in the time/wild pig captured among traps. Our findings demonstrate that when operated by wildlife professionals, multiple designs of wild pig traps can be effective at removing targeted groups of wild pigs. Further, our results offer insights regarding the performance of wild pig trap types across seasons that can be used to inform wild pig trapping programs.

Taylor, Charles R. [Warnell School of Forestry and↗