Surviving the cold: molecular analyses of insect cryoprotective dehydration in the Arctic springtail Megaphorura arctica (Tullberg)

<p>Abstract</p> <p>Background</p> <p>Insects provide tractable models for enhancing our understanding of the physiological and cellular processes that enable survival at extreme low temperatures. They possess three main strategies to survive the cold: freeze tolerance, freeze avoidance or cryoprotective dehydration, of which the latter method is exploited by our model species, the Arctic springtail <it>Megaphorura arctica</it>, formerly <it>Onychiurus arcticus </it>(Tullberg 1876). The physiological mechanisms underlying cryoprotective dehydration have been well characterised in <it>M. arctica </it>and to date this process has been described in only a few other species: the Antarctic nematode <it>Panagrolaimus davidi</it>, an enchytraied worm, the larvae of the Antarctic midge <it>Belgica antarctica </it>and the cocoons of the earthworm <it>Dendrobaena octaedra</it>. There are no in-depth molecular studies on the underlying cold survival mechanisms in any species.</p> <p>Results</p> <p>A cDNA microarray was generated using 6,912 <it>M. arctica </it>clones printed in duplicate. Analysis of clones up-regulated during dehydration procedures (using both cold- and salt-induced dehydration) has identified a number of significant cellular processes, namely the production and mobilisation of trehalose, protection of cellular systems via small heat shock proteins and tissue/cellular remodelling during the dehydration process. Energy production, initiation of protein translation and cell division, plus potential tissue repair processes dominate genes identified during recovery. Heat map analysis identified a duplication of the trehalose-6-phosphate synthase (TPS) gene in <it>M. arctica </it>and also 53 clones co-regulated with TPS, including a number of membrane associated and cell signalling proteins. Q-PCR on selected candidate genes has also contributed to our understanding with glutathione-S-transferase identified as the major antioxdidant enzyme protecting the cells during these stressful procedures, and a number of protein kinase signalling molecules involved in recovery.</p> <p>Conclusion</p> <p>Microarray analysis has proved to be a powerful technique for understanding the processes and genes involved in cryoprotective dehydration, beyond the few candidate genes identified in the current literature. Dehydration is associated with the mobilisation of trehalose, cell protection and tissue remodelling. Energy production, leading to protein production, and cell division characterise the recovery process. Novel membrane proteins, along with aquaporins and desaturases, have been identified as promising candidates for future functional analyses to better understand membrane remodelling during cellular dehydration.</p

​Rapid response to experimental warming of a microbial community inhabiting High Arctic patterned ground soil

The influence of climate change on microbial communities inhabiting the sparsely vegetated patterned ground soils that are widespread across the High Arctic is poorly understood. Here, in a four-year experiment on Svalbard, we warmed patterned ground soil with open top chambers and biannually irrigated the soil to predict the responses of its microbial community to rising temperatures and precipitation. A 1 °C rise in summertime soil temperature caused 44% and 78% increases in CO2 efflux and CH4 consumption, respectively, and a 32% increase in the frequency of bacterial 16S ribosomal RNA genes. Bacterial alpha diversity was unaffected by the treatments, but, of the 40 most frequent bacterial taxa, warming caused 44–45% reductions in the relative abundances of a Sphingomonas sp. and Ferruginibacter sp. and 33–91% increases in those of a Phenylobacterium sp. and a member of the Acetobacteraceae. Warming did not influence the frequency of fungal internal transcribed spacer 2 copies, and irrigation had no effects on the measured variables. Our study suggests rapid changes to the activities and abundances of microbes, and particularly bacteria, in High Arctic patterned ground soils as they warm. At current rates of soil warming on Svalbard (0.8 °C per decade), we anticipate that similar effects to those reported here will manifest themselves in the natural environment by approximately the mid 2030s

Nicotine enhances an auditory Event-Related Potential component which is inversely related to habituation

Nicotine is a psychoactive substance that is commonly consumed in the context of music. However, the reason why music and nicotine are coconsumed is uncertain. One possibility is that nicotine affects cognitive processes relevant to aspects of music appreciation in a beneficial way. Here we investigated this possibility using Event-Related Potentials (ERPs). Participants underwent a simple decision-making task (to maintain attentional focus), responses to which were signaled by auditory stimuli. Unlike most previous research looking at the effects of nicotine on auditory processing, we used tones of different pitch, a fundamental element of music. In addition, unlike most other studies, we tested non-smoking subjects to avoid withdrawal-related complications. We found that nicotine (4.0 mg, administered as gum) increased P2 amplitude in the frontal region. Since a decrease in P2 amplitude and latency is related to habituation processes, and an enhanced ability to disengage from irrelevant stimuli, our findings suggest that nicotine may cause a reduction in habituation, resulting in non-smokers being less able to adapt to repeated stimuli. A corollary of that decrease in adaptation may be that nicotine extends the temporal window during which a listener is able and willing to engage with a piece of music

Pure phase-locking of beta/gamma oscillation contributes to the N30 frontal component of somatosensory evoked potentials

BACKGROUND: Evoked potentials have been proposed to result from phase-locking of electroencephalographic (EEG) activities within specific frequency bands. However, the respective contribution of phasic activity and phase resetting of ongoing EEG oscillation remains largely debated. We here applied the EEGlab procedure in order to quantify the contribution of electroencephalographic oscillation in the generation of the frontal N30 component of the somatosensory evoked potentials (SEP) triggered by median nerve electrical stimulation at the wrist. Power spectrum and intertrial coherence analysis were performed on EEG recordings in relation to median nerve stimulation. RESULTS: The frontal N30 component was accompanied by a significant phase-locking of beta/gamma oscillation (25-35 Hz) and to a lesser extent of 80 Hz oscillation. After the selection in each subject of the trials for which the power spectrum amplitude remained unchanged, we found pure phase-locking of beta/gamma oscillation (25-35 Hz) peaking about 30 ms after the stimulation. Transition across trials from uniform to normal phase distribution revealed temporal phase reorganization of ongoing 30 Hz EEG oscillations in relation to stimulation. In a proportion of trials, this phase-locking was accompanied by a spectral power increase peaking in the 30 Hz frequency band. This corresponds to the complex situation of 'phase-locking with enhancement' in which the distinction between the contribution of phasic neural event versus EEG phase resetting is hazardous. CONCLUSION: The identification of a pure phase-locking in a large proportion of the SEP trials reinforces the contribution of the oscillatory model for the physiological correlates of the frontal N30. This may imply that ongoing EEG rhythms, such as beta/gamma oscillation, are involved in somatosensory information processing.Comparative StudyJournal ArticleResearch Support, Non-U.S. Gov'tinfo:eu-repo/semantics/publishe

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

RNA preservation of Antarctic marine invertebrates

Fifteen species of marine invertebrate commonly occurring in the near-shore environment of Rothera base, Antarctica, were used to test tissue sample storage protocols with regard to preservation of RNA integrity. After animal collection, the tissues were either immediately extracted for RNA or stored at -80C after having been, either directly flash frozen in liquid nitrogen or preserved in a commercial RNA storage solution, for extraction in the UK. In four cases, direct flash freezing produced enhanced RNA integrity compared with samples in the commercial storage solution. A subset of samples were further tested for the preferred temperature of storage in the commercial reagent. RNA integrity was well preserved at both ?4 and -20C over periods of 2 months, but degradation was rapid in tissues stored at room temperature. Eight out of the fifteen species only produced a single ribosomal band on gel electrophoresis. This survey provides a guide for tissue transport of Polar cold water marine invertebrates. Keywords Tissue preservation Tissue transport 28 s ribosomal RNA Echinoderms Molluscs Introduction RNA preservation is sometimes problematic in non-model species but this is particularly the case when dealing with environmental species. Logistical issues often surround the ability to effectively preserve field-collected samples for RNA analyses. Whilst rapid flash freezing in liquid nitrogen generally solves this problem, it is not often available because of the remote nature of the work. Even when such a facility is available on site at a field station, it usually cannot be transported to the actual, more remote specimen collection site. Also, -80C storage may not be possible during transportation from the field site to the main research institute, often thousands of miles away. Antarctic specimens have the additional issue of operating at temperatures that most species would consider cold and hence cool stow is less effective at reducing tissue degradation than with, for example, those taken from mammalian species. Hence, we decided to carry out a study of effective storage protocols for the most common invertebrates found in the near-shore marine environment in Marguerite Bay close to Rothera research station, Antarctica

Gene expression associated with changes in cold tolerance levels of the Antarctic springtail, Cryptopygus antarcticus

The ability of the Antarctic microarthropod Cryptopygus antarcticus (Collembola, Isotomidae) to survive low temperatures has been well studied at the physiological level, with recent investigations indicating the importance of the moulting process in conferring this ability. This study investigated gene expression in groups of C. antarcticus that have distinct differences in their ability to survive low temperatures. A microarray containing c. 5400 C. antarcticus expressed sequence tags was used to investigate gene expression differences between groups of animals with different supercooling points (SCP), and to low temperatures close to their SCP. By demonstrating the involvement of moult-related genes in the differential survival of two groups of C. antarcticus with distinct SCP profiles, the results of this investigation add support to the suggestion that moulting plays a role in conferring cold tolerance in C. antarcticus

Transcriptional response to heat stress in the Antarctic bivalve Laternula elliptica

Our ability to predict animal responses to temperature changes is currently limited and more sensitive methods of identifying mechanisms, limits and thresholds are required. Antarctic marine ectotherms are excellent candidates for the study of warming seawater temperatures as they represent some of the most stenothermal species on Earth. They live within a narrow thermal window and exhibit reduced physiological capacities with small elevations in temperature. Whilst the physiological effects of elevated temperatures are well documented in these species, knowledge at the molecular level is scant and hence our understanding of the cellular response is still limited. To increase our knowledge in this area, a cDNA microarray consisting of 8448 clones was constructed to study the heat stress response of Laternula elliptica, an infaunal Antarctic bivalve. Eight clams were exposed to 12 h heat stress at 3 degrees C and their gene expression profiles were individually compared to a pooled reference of non heat shocked clams. Significant changes in the expression of 294 clones, representing 160 transcripts were observed. Of these, 33 were identified by sequence similarity searches and classified to a variety of cellular functions including protein turnover, folding and chaperoning, intracellular signalling and trafficking and cytoskeletal activity. These results suggest that antioxidant genes and genes involved in Ca2+ signalling and homeostasis represent potential biomarkers as to the physiological state of this species under thermal stress. (C) 2010 Elsevier B.V. All rights reserved

In Search of Covariates of HIV-1 Subtype B Spread in the United States-A Cautionary Tale of Large-Scale Bayesian Phylogeography.

Infections with HIV-1 group M subtype B viruses account for the majority of the HIV epidemic in the Western world. Phylogeographic studies have placed the introduction of subtype B in the United States in New York around 1970, where it grew into a major source of spread. Currently, it is estimated that over one million people are living with HIV in the US and that most are infected with subtype B variants. Here, we aim to identify the drivers of HIV-1 subtype B dispersal in the United States by analyzing a collection of 23,588 pol sequences, collected for drug resistance testing from 45 states during 2004-2011. To this end, we introduce a workflow to reduce this large collection of data to more computationally-manageable sample sizes and apply the BEAST framework to test which covariates associate with the spread of HIV-1 across state borders. Our results show that we are able to consistently identify certain predictors of spread under reasonable run times across datasets of up to 10,000 sequences. However, the general lack of phylogenetic structure and the high uncertainty associated with HIV trees make it difficult to interpret the epidemiological relevance of the drivers of spread we are able to identify. While the workflow we present here could be applied to other virus datasets of a similar scale, the characteristic star-like shape of HIV-1 phylogenies poses a serious obstacle to reconstructing a detailed evolutionary and spatial history for HIV-1 subtype B in the US