Transcriptional response to heat stress in the Antarctic bivalve Laternula elliptica

Abstract 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 °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.

[1]  G. Somero,et al.  Temperature Tolerance of Some Antarctic Fishes , 1967, Science.

[2]  Jonathon H Stillman,et al.  A cDNA microarray analysis of the response to heat stress in hepatopancreas tissue of the porcelain crab Petrolisthes cinctipes. , 2007, Comparative biochemistry and physiology. Part D, Genomics & proteomics.

[3]  Y. Tsay,et al.  Ribosomal protein synthesis is not regulated at the translational level in Saccharomyces cerevisiae: balanced accumulation of ribosomal proteins L16 and rp59 is mediated by turnover of excess protein. , 1988, Genes & development.

[4]  I. Johnston,et al.  Evolution and adaptive radiation of antarctic fishes. , 1996, Trends in ecology & evolution.

[5]  Lloyd S. Peck,et al.  Extreme sensitivity of biological function to temperature in Antarctic marine species , 2004 .

[6]  R. Kleppe,et al.  Does isoform diversity explain functional differences in the 14-3-3 protein family? , 2006, Current pharmaceutical biotechnology.

[7]  I. Ahn,et al.  Expression of heat shock protein 70 in the thermally stressed Antarctic clam Laternula elliptica , 2007, Cell stress & chaperones.

[8]  I. Ahn,et al.  Analysis of ESTs and expression of two peroxiredoxins in the thermally stressed Antarctic bivalve Laternula elliptica. , 2008, Fish & shellfish immunology.

[9]  H. Pörtner,et al.  Climate change and temperature-dependent biogeography: oxygen limitation of thermal tolerance in animals , 2001, Naturwissenschaften.

[10]  R. Crozier,et al.  Heterologous microarray experiments used to identify the early gene response to heat stress in a coral reef fish , 2007, Molecular ecology.

[11]  Melody S Clark,et al.  Insights into shell deposition in the Antarctic bivalve Laternula elliptica: gene discovery in the mantle transcriptome using 454 pyrosequencing , 2010, BMC Genomics.

[12]  H. Pörtner,et al.  Production of reactive oxygen species by isolated mitochondria of the Antarctic bivalve Laternula elliptica (King and Broderip) under heat stress. , 2003, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[13]  Gordon K. Smyth,et al.  A comparison of background correction methods for two-colour microarrays , 2007, Bioinform..

[14]  Cathy H. Wu,et al.  The Universal Protein Resource (UniProt) , 2004, Nucleic Acids Res..

[15]  The simultaneous immunological detection of four stress-70 protein isoforms in Mytilus edulis , 1995 .

[16]  J. Figueroa,et al.  Environmental acclimatization of the carp modulates the transcription of β‐actin , 2001 .

[17]  F. Morón,et al.  Genetic Structure of the Spanish Population , 2010, BMC Genomics.

[18]  E. Davidson,et al.  Experimentally based sea urchin gene regulatory network and the causal explanation of developmental phenomenology , 2009, Wiley interdisciplinary reviews. Systems biology and medicine.

[19]  D. Kültz,et al.  Molecular and evolutionary basis of the cellular stress response. , 2005, Annual review of physiology.

[20]  Matthias Mann,et al.  Analysis of Nucleolar Protein Dynamics Reveals the Nuclear Degradation of Ribosomal Proteins , 2007, Current Biology.

[21]  G. Somero,et al.  Changes in gene expression associated with acclimation to constant temperatures and fluctuating daily temperatures in an annual killifish Austrofundulus limnaeus , 2004, Journal of Experimental Biology.

[22]  H. Pörtner,et al.  Climate variations and the physiological basis of temperature dependent biogeography: systemic to molecular hierarchy of thermal tolerance in animals. , 2002, Comparative biochemistry and physiology. Part A, Molecular & integrative physiology.

[23]  F. Hartl,et al.  Structure of the Molecular Chaperone Prefoldin Unique Interaction of Multiple Coiled Coil Tentacles with Unfolded Proteins , 2000, Cell.

[24]  J. Vandekerckhove,et al.  Prefoldin, a Chaperone that Delivers Unfolded Proteins to Cytosolic Chaperonin , 1998, Cell.

[25]  A. Sobieszek,et al.  Effect of actin C-terminal modification on tropomyosin isoforms binding and thin filament regulation , 2009, Biochimica et biophysica acta.

[26]  S. Müller,et al.  The double life of HMGB1 chromatin protein: architectural factor and extracellular signal , 2001 .

[27]  L. Peck,et al.  Metabolic Demand, Oxygen Supply, and Critical Temperatures in the Antarctic Bivalve Laternula elliptica , 2002, Physiological and Biochemical Zoology.

[28]  S. Bagrodia,et al.  Pak to the future. , 1999, Trends in cell biology.

[29]  S Rozen,et al.  Primer3 on the WWW for general users and for biologist programmers. , 2000, Methods in molecular biology.

[30]  A. Krogh,et al.  The Respiratory Exchange of Animals and Man , 2009, Nature.

[31]  Gordon K Smyth,et al.  Linear Models and Empirical Bayes Methods for Assessing Differential Expression in Microarray Experiments , 2004, Statistical applications in genetics and molecular biology.

[32]  Rafael A. Irizarry,et al.  Bioinformatics and Computational Biology Solutions using R and Bioconductor , 2005 .

[33]  M. Berridge,et al.  Calcium signalling--an overview. , 2001, Seminars in cell & developmental biology.

[34]  G. Somero,et al.  Thermal limits and adaptation in marine Antarctic ectotherms: an integrative view , 2007, Philosophical Transactions of the Royal Society B: Biological Sciences.

[35]  Gordon K. Smyth,et al.  limma: Linear Models for Microarray Data , 2005 .

[36]  G. Somero,et al.  The cellular response to heat stress in the goby Gillichthys mirabilis: a cDNA microarray and protein-level analysis , 2006, Journal of Experimental Biology.

[37]  P. Leung,et al.  The recovery of some components of the renin angiotensin system in the rat pancreas after chronic exposure to hypoxic condition. , 2003, Journal of molecular endocrinology.

[38]  Terry Speed,et al.  Normalization of cDNA microarray data. , 2003, Methods.

[39]  Zhanjiang Liu,et al.  Differential gene expression in the brain of channel catfish (Ictalurus punctatus) in response to cold acclimation , 2002, Molecular Genetics and Genomics.

[40]  G. Chainy,et al.  Biochemical markers of oxidative stress in Perna viridis exposed to mercury and temperature. , 2007, Chemico-biological interactions.

[41]  Gordon K. Smyth,et al.  Use of within-array replicate spots for assessing differential expression in microarray experiments , 2005, Bioinform..

[42]  Melody S Clark,et al.  HSP70 heat shock proteins and environmental stress in Antarctic marine organisms: A mini-review. , 2009, Marine genomics.

[43]  G. Burns,et al.  Cold hardening processes in the Antarctic springtail, Cryptopygus antarcticus: clues from a microarray. , 2008, Journal of insect physiology.

[44]  Jonathon H Stillman,et al.  Acclimation Capacity Underlies Susceptibility to Climate Change , 2003, Science.

[45]  Richard T. Di Giulio,et al.  Prooxidant and antioxidant mechanisms in aquatic organisms , 1991 .

[46]  Manuel Manchado,et al.  Comparative sequence analysis of the complete set of 40S ribosomal proteins in the Senegalese sole (Solea senegalensis Kaup) and Atlantic halibut (Hippoglossus hippoglossus L.) (Teleostei: Pleuronectiformes): phylogeny and tissue- and development-specific expression , 2007, BMC Evolutionary Biology.

[47]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[48]  S. Sunagawa,et al.  Differential gene expression during thermal stress and bleaching in the Caribbean coral Montastraea faveolata , 2008, Molecular ecology.

[49]  Michael O'Donnell,et al.  Gene expression in the intertidal mussel Mytilus californianus: physiological response to environmental factors on a biogeographic scale , 2008 .

[50]  L. Peck,et al.  Hyperoxia alleviates thermal stress in the Antarctic bivalve, Laternula elliptica: evidence for oxygen limited thermal tolerance , 2006, Polar Biology.

[51]  Lloyd S. Peck,et al.  Antarctic marine molluscs do have an HSP70 heat shock response , 2008, Cell Stress and Chaperones.

[52]  I. Ahn,et al.  Response of antioxidant defence systems to thermal stress in the Antarctic clam Laternula elliptica , 2008, Antarctic Science.

[53]  G. Somero,et al.  Biochemical Adaptation: Mechanism and Process in Physiological Evolution , 1984 .

[54]  Lloyd S. Peck,et al.  Animal temperature limits and ecological relevance: effects of size, activity and rates of change , 2009 .

[55]  H. Pörtner,et al.  Oxygen limitation of thermal tolerance defined by cardiac and ventilatory performance in spider crab, Maja squinado. , 2000, American journal of physiology. Regulatory, integrative and comparative physiology.

[56]  M. Ashburner,et al.  Gene Ontology: tool for the unification of biology , 2000, Nature Genetics.

[57]  R. Angerer,et al.  Gene Regulatory Network Interactions in Sea Urchin Endomesoderm Induction , 2009, PLoS biology.

[58]  J. O. Thomas,et al.  HMG1 and 2: architectural DNA-binding proteins. , 2001, Biochemical Society transactions.

[59]  M. Feder,et al.  Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. , 1999, Annual review of physiology.

[60]  G. Somero,et al.  cDNA microarray analysis reveals the capacity of the cold-adapted Antarctic fish Trematomus bernacchii to alter gene expression in response to heat stress , 2009, Polar Biology.

[61]  G. Horgan,et al.  Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR , 2002 .

[62]  N. Segev,et al.  Conservation of the TRAPPII-specific subunits of a Ypt/Rab exchanger complex , 2007, BMC Evolutionary Biology.

[63]  G. Somero,et al.  Time Course and Magnitude of Synthesis of Heat‐Shock Proteins in Congeneric Marine Snails (Genus Tegula) from Different Tidal Heights , 2000, Physiological and Biochemical Zoology.

[64]  H. Ellegren.  Sequencing goes 454 and takes large‐scale genomics into the wild , 2008, Molecular ecology.

[65]  S. Ebashi,et al.  Calcium ion and muscle contraction. , 1968, Progress in biophysics and molecular biology.

[66]  G. Somero,et al.  The threshold induction temperature of the 90-kDa heat shock protein is subject to acclimatization in eurythermal goby fishes (genus Gillichthys). , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[67]  E. C. Toescu.  Hypoxia sensing and pathways of cytosolic Ca2+ increases. , 2004, Cell calcium.

[68]  A. Viarengo,et al.  Exposure to elevated temperatures and hydrogen peroxide elicits oxidative stress and antioxidant response in the Antarctic intertidal limpet Nacella concinna , 1998 .

[69]  M. Lindström.  Emerging functions of ribosomal proteins in gene-specific transcription and translation. , 2009, Biochemical and biophysical research communications.

[70]  Y. Takahashi.  The 14-3-3 Proteins: Gene, Gene Expression, and Function , 2003, Neurochemical Research.

[71]  S. Lindquist,et al.  The function of heat-shock proteins in stress tolerance: degradation and reactivation of damaged proteins. , 1993, Annual review of genetics.

[72]  V. P. Collins,et al.  Global amplification of mRNA by template-switching PCR: linearity and application to microarray analysis. , 2003, Nucleic acids research.

[73]  John Quackenbush,et al.  TIGR Gene Indices clustering tools (TGICL): a software system for fast clustering of large EST datasets , 2003, Bioinform..