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Rhythms of gene expression in mussel Mytilus galloprovincialis during an annual cycle
Abstracts / Comparative Biochemistry and Physiology, Part A 154 (2009) S1–S6
months of treatment. The stress induced by Diuron changes the percentage of muscle carcase, spleen and liver while the stress by handle affects the percentage of head, viscera, gut, perivisceral fat, condition factor, weight, length, high, head length with a general reduction of growth in handle stress fish. doi:10.1016/j.cbpa.2009.05.026
13. Metal-dependent transcription of Metallothionein Isoform genes in the terrestrial snail Cantareus aspersus
S5
tions. The hierarchical clustering of mussel digestive gland gene expression profiles show similar trends in months (May, June and July), (August, September and October), (November and December), (Febrary, March and April). Genes involved in chitin metabolism (Chitinase and Chitotriosidase) showed a very a marked trend with a higher expression during summer. The acrosomal major pritein M3 is found to be up regulated during October. Moreover, the differential gene expression profile of females respect to male was also assessed in animals during the four stages of the mussel gonad development. Data show a marked divergent profile during stage II (developing) and stage III (ripe).
doi:10.1016/j.cbpa.2009.05.028 M. Höckner, D. Schuler, K. Stefanon, R. Fantur (University of Innsbruck, Austria); A. de Vaufleury (University of Franche-Comté, Besançon, France); R. Dallinger (University of Innsbruck, Austria) Detoxification of heavy metals on the one hand and homeostasis of essential trace elements on the other hand are necessities every organism has to deal with. In particular, Metallothioneins (MTs) are proteins which bear the potential to fulfil these contradictive requirements. In terrestrial gastropods, different MT isoforms overtake these diverse functions. The terrestrial helicid snail Cantareus aspersus, for example, possesses three MT isoforms: Two of these are strictly metal-specific (a Cd- and a Cu-specific isoform), while a third intermediate isoforms binds both metals, Cd and Cu. All three isoforms were identified at the cDNA level. In addition, the corresponding promotor regions were sequenced and analysed with respect to relevant transcription factor binding sites. Transcriptional activation of all three isoforms upon exposure to cadmium and copper over a period of 29 days was assessed by real-time detection PCR. In addition, respective metal concentrations in snail tissues were quantified during the exposure period. Transcriptional induction after Cd exposure was only observed for the Cd-MT gene, whereas copper had no effect on transcriptional activation for any of the three isoform genes. This leads to the conclusion that the Cd-MT gene must be involved in Cd detoxification. On the other hand, the Cu- and Cd/CuMT isoform genes may play a role in Cu homeostasis. The differing transcriptional activation patterns observed in the three isoform genes may be explained by differences in promotor structure and particularly, by different sequence and organisation of metal responsive elements (MREs) located on specific promoter regions of the three genes (supported by the Austrian Science Foundation, project No. P19782-B03). doi:10.1016/j.cbpa.2009.05.027
14. Rhythms of gene expression in mussel Mytilus galloprovincialis during an annual cycle A. Negri, A. Viarengo, F. Dondero (University of Piemonte Orientale “Avogadro”, Italy), M. Banni (Institut Supérieur Agronomique, Tunisia); F. Mignone (University of Milan, Italy) The marine mussel Mytilus galloprovincialis is an intertidal filterfeeders commonly used as biosensors of coastal pollution. Environmental changes due to seasonal variation (temperature, oxygen tension, nutrient concentration, etc.) deeply affect the molecular response of mussels. This study was undertaken to investigate the seasonal variation of digestive gland and mantel relative transcript abundance in males and female individuals of the marine mussel Mytilus galloprovincialis (LAM). A medium density microarray (1.7 K probes) was used in dual color cDNA probes competitive hybridiza-
15. Xenobiotic detoxification in Australian marsupials – An overview S.N.T. Ngo (Charles Darwin University, Australia); I. Stupans, R.A. McKinnon (University of South Australia, Australia) Like humans and other mammals, Australian marsupials are constantly exposed to a diverse array of xenobiotics that come from a wide variety of sources, including natural toxins in the diet, industrial and agricultural contaminants, and other environmental exposures. Enzymatic modification of xenobiotics can potentially alter their activity and toxicity. Knowledge of these processes is vital to understanding the consequences of exposure to xenobiotics whether this is from the xenobiotic itself or from its metabolites. To date, extensive studies of xenobiotic detoxification have been undertaken in eutherian mammals, primarily humans, mice and rats. In sharp contrast to the plethora of studies in eutherian mammals, few studies have been undertaken in marsupials. The current study provides an overview of research being undertaken over several years that has provided unique insights into xenobiotic detoxification in Australian marsupials, including studies from our laboratories. The focus of this study is on enzymatic data on phase I and phase II metabolisms, the multiplicity of detoxifying enzymes/pathways involved, and induction/inhibition of CYPs by dietary Eucalyptus terpenes. The study also highlights the important interspecies differences among these unique animals. The results of preliminary studies on marsupial detoxification demonstrated that, relative to eutherian species such as humans and rats, marsupials display several significant differences in xenobiotic metabolising enzymes at the biochemical level. Further investigations into enzymes concerning dietary Eucalyptus terpene detoxification in marsupials are critically important in identifying the significances of these preliminary observations and in understanding the adaptation of marsupial herbivores to their unique Eucalyptus diet.
doi:10.1016/j.cbpa.2009.05.029
16. Characterization and expression analysis of bacteria challenged calmodulin homologue in disk abalone C. Nikapitiya, J. Lee (Jeju National University, Republic of Korea) Calcium ion plays a crucial role in the various biochemical processes of metabolism and physiology in all the eukaryotics. Transient increases in calcium concentrations with in the cell occur due to various stresses including pathogenesis and cells have developed multitude ways to control and regulate the cellular processes. Calmodulin is a calcium binding protein, which transducers
months of treatment. The stress induced by Diuron changes the percentage of muscle carcase, spleen and liver while the stress by handle affects the percentage of head, viscera, gut, perivisceral fat, condition factor, weight, length, high, head length with a general reduction of growth in handle stress fish. doi:10.1016/j.cbpa.2009.05.026
13. Metal-dependent transcription of Metallothionein Isoform genes in the terrestrial snail Cantareus aspersus
S5
tions. The hierarchical clustering of mussel digestive gland gene expression profiles show similar trends in months (May, June and July), (August, September and October), (November and December), (Febrary, March and April). Genes involved in chitin metabolism (Chitinase and Chitotriosidase) showed a very a marked trend with a higher expression during summer. The acrosomal major pritein M3 is found to be up regulated during October. Moreover, the differential gene expression profile of females respect to male was also assessed in animals during the four stages of the mussel gonad development. Data show a marked divergent profile during stage II (developing) and stage III (ripe).
doi:10.1016/j.cbpa.2009.05.028 M. Höckner, D. Schuler, K. Stefanon, R. Fantur (University of Innsbruck, Austria); A. de Vaufleury (University of Franche-Comté, Besançon, France); R. Dallinger (University of Innsbruck, Austria) Detoxification of heavy metals on the one hand and homeostasis of essential trace elements on the other hand are necessities every organism has to deal with. In particular, Metallothioneins (MTs) are proteins which bear the potential to fulfil these contradictive requirements. In terrestrial gastropods, different MT isoforms overtake these diverse functions. The terrestrial helicid snail Cantareus aspersus, for example, possesses three MT isoforms: Two of these are strictly metal-specific (a Cd- and a Cu-specific isoform), while a third intermediate isoforms binds both metals, Cd and Cu. All three isoforms were identified at the cDNA level. In addition, the corresponding promotor regions were sequenced and analysed with respect to relevant transcription factor binding sites. Transcriptional activation of all three isoforms upon exposure to cadmium and copper over a period of 29 days was assessed by real-time detection PCR. In addition, respective metal concentrations in snail tissues were quantified during the exposure period. Transcriptional induction after Cd exposure was only observed for the Cd-MT gene, whereas copper had no effect on transcriptional activation for any of the three isoform genes. This leads to the conclusion that the Cd-MT gene must be involved in Cd detoxification. On the other hand, the Cu- and Cd/CuMT isoform genes may play a role in Cu homeostasis. The differing transcriptional activation patterns observed in the three isoform genes may be explained by differences in promotor structure and particularly, by different sequence and organisation of metal responsive elements (MREs) located on specific promoter regions of the three genes (supported by the Austrian Science Foundation, project No. P19782-B03). doi:10.1016/j.cbpa.2009.05.027
14. Rhythms of gene expression in mussel Mytilus galloprovincialis during an annual cycle A. Negri, A. Viarengo, F. Dondero (University of Piemonte Orientale “Avogadro”, Italy), M. Banni (Institut Supérieur Agronomique, Tunisia); F. Mignone (University of Milan, Italy) The marine mussel Mytilus galloprovincialis is an intertidal filterfeeders commonly used as biosensors of coastal pollution. Environmental changes due to seasonal variation (temperature, oxygen tension, nutrient concentration, etc.) deeply affect the molecular response of mussels. This study was undertaken to investigate the seasonal variation of digestive gland and mantel relative transcript abundance in males and female individuals of the marine mussel Mytilus galloprovincialis (LAM). A medium density microarray (1.7 K probes) was used in dual color cDNA probes competitive hybridiza-
15. Xenobiotic detoxification in Australian marsupials – An overview S.N.T. Ngo (Charles Darwin University, Australia); I. Stupans, R.A. McKinnon (University of South Australia, Australia) Like humans and other mammals, Australian marsupials are constantly exposed to a diverse array of xenobiotics that come from a wide variety of sources, including natural toxins in the diet, industrial and agricultural contaminants, and other environmental exposures. Enzymatic modification of xenobiotics can potentially alter their activity and toxicity. Knowledge of these processes is vital to understanding the consequences of exposure to xenobiotics whether this is from the xenobiotic itself or from its metabolites. To date, extensive studies of xenobiotic detoxification have been undertaken in eutherian mammals, primarily humans, mice and rats. In sharp contrast to the plethora of studies in eutherian mammals, few studies have been undertaken in marsupials. The current study provides an overview of research being undertaken over several years that has provided unique insights into xenobiotic detoxification in Australian marsupials, including studies from our laboratories. The focus of this study is on enzymatic data on phase I and phase II metabolisms, the multiplicity of detoxifying enzymes/pathways involved, and induction/inhibition of CYPs by dietary Eucalyptus terpenes. The study also highlights the important interspecies differences among these unique animals. The results of preliminary studies on marsupial detoxification demonstrated that, relative to eutherian species such as humans and rats, marsupials display several significant differences in xenobiotic metabolising enzymes at the biochemical level. Further investigations into enzymes concerning dietary Eucalyptus terpene detoxification in marsupials are critically important in identifying the significances of these preliminary observations and in understanding the adaptation of marsupial herbivores to their unique Eucalyptus diet.
doi:10.1016/j.cbpa.2009.05.029
16. Characterization and expression analysis of bacteria challenged calmodulin homologue in disk abalone C. Nikapitiya, J. Lee (Jeju National University, Republic of Korea) Calcium ion plays a crucial role in the various biochemical processes of metabolism and physiology in all the eukaryotics. Transient increases in calcium concentrations with in the cell occur due to various stresses including pathogenesis and cells have developed multitude ways to control and regulate the cellular processes. Calmodulin is a calcium binding protein, which transducers