Characterization of prostaglandins in the Pacific oyster  Crassostrea gigas : evidence for a role in immune response  Mack...
Molecules with Diverse Functions <ul><li>Vertebrates </li></ul><ul><ul><li>Mediate swelling and inflammation  </li></ul></...
Molecules with Diverse Functions <ul><ul><li>Invertebrates </li></ul></ul><ul><ul><ul><li>Ion transport  </li></ul></ul></...
What are prostaglandins? <ul><li>Eicosanoids: a family of biologically active metabolites of C20 polyunsaturated fatty aci...
Arachidonic Acid Cascade PGE receptor
Outline <ul><li>Characterize molecules in prostaglandin pathway </li></ul><ul><ul><li>ID gene fragments using ESTs </li></...
Arachidonic Acid Cascade PGE receptor
Sequences PGE receptor ESTs: snail, mussel <ul><li>ESTs: oyster!,mussel </li></ul><ul><li>AM856036 </li></ul>ESTs: ongoing...
Data PGE receptor
Full-length Sequence PGE receptor <ul><li>STATS : </li></ul><ul><li>Size: 1700 bp </li></ul><ul><li>pred. # introns: 10 </...
COX Gene Similarities
Outline <ul><li>Characterize molecules in prostaglandin pathway </li></ul><ul><ul><li>ID gene fragments from genomics data...
Experiments <ul><li>Vibrio tubiashii  exposures  </li></ul><ul><li>whole organism   plated    hemocytes </li></ul>
Analytical Results <ul><li>PGE (ELISA) </li></ul><ul><li>Receptor (PCR) </li></ul>PGE receptor
Vibrio exposure-ELISA results PGE conjugate PGE sample 100% 0%
Gill Tissue/Vibrio– qPCR PGE receptor
Plated hemocytes/Vibrio - qPCR
Outline <ul><li>What are prostaglandins? </li></ul><ul><li>Characterization of genes involved in biosynthesis of prostagla...
Applied Research <ul><li>Physiological Processes: </li></ul><ul><li>Immune Response </li></ul><ul><li>Ion transport </li><...
Applied Research <ul><li>Prostaglandins appear to play a role in the immune response of oysters. </li></ul><ul><li>In vitr...
Acknowledgements <ul><li>Advisor, Steven Roberts </li></ul><ul><li>Research scientist, Sam White </li></ul><ul><li>Friedma...
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SAFS GSS Nov 2008: Characterization of prostaglandins in the Pacific oyster Crassostrea gigas: evidence for a role in immune response

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SAFS GSS Nov 2008: Characterization of prostaglandins in the Pacific oyster Crassostrea gigas: evidence for a role in immune response

  1. 1. Characterization of prostaglandins in the Pacific oyster Crassostrea gigas : evidence for a role in immune response Mackenzie Gavery, M.S. Student Roberts Lab
  2. 2. Molecules with Diverse Functions <ul><li>Vertebrates </li></ul><ul><ul><li>Mediate swelling and inflammation </li></ul></ul><ul><ul><ul><li>(COX inhibitors) </li></ul></ul></ul><ul><ul><li>Inducing fever </li></ul></ul><ul><ul><li>Maintaining pregnancy/reproductive timing </li></ul></ul><ul><ul><li>Regulating ion transport </li></ul></ul>
  3. 3. Molecules with Diverse Functions <ul><ul><li>Invertebrates </li></ul></ul><ul><ul><ul><li>Ion transport </li></ul></ul></ul><ul><ul><ul><li>Reproduction </li></ul></ul></ul><ul><ul><ul><li>Immune Response </li></ul></ul></ul><ul><ul><ul><ul><li>recently discovered a gene encoding prostaglandin E 2 receptor in Pacific oysters </li></ul></ul></ul></ul><ul><ul><ul><ul><li>significantly upregulated after exposure to Vibrio tubiashii </li></ul></ul></ul></ul>Roberts et al
  4. 4. What are prostaglandins? <ul><li>Eicosanoids: a family of biologically active metabolites of C20 polyunsaturated fatty acids. </li></ul><ul><li>Prostaglandins biosynthesis begins with the from arachidonic acid, an omega-6 fatty acid. </li></ul><ul><li>prostaglandin E2: </li></ul>
  5. 5. Arachidonic Acid Cascade PGE receptor
  6. 6. Outline <ul><li>Characterize molecules in prostaglandin pathway </li></ul><ul><ul><li>ID gene fragments using ESTs </li></ul></ul><ul><ul><li>Tissue distribution of oyster </li></ul></ul><ul><ul><li>Isolate complete ORF of oyster COX gene </li></ul></ul><ul><li>Challenge Experiments </li></ul><ul><li>Applied research </li></ul>
  7. 7. Arachidonic Acid Cascade PGE receptor
  8. 8. Sequences PGE receptor ESTs: snail, mussel <ul><li>ESTs: oyster!,mussel </li></ul><ul><li>AM856036 </li></ul>ESTs: ongoing <ul><li>ESTs: </li></ul><ul><li>EW777722 </li></ul>
  9. 9. Data PGE receptor
  10. 10. Full-length Sequence PGE receptor <ul><li>STATS : </li></ul><ul><li>Size: 1700 bp </li></ul><ul><li>pred. # introns: 10 </li></ul><ul><li>Amino acids: 587 </li></ul>
  11. 11. COX Gene Similarities
  12. 12. Outline <ul><li>Characterize molecules in prostaglandin pathway </li></ul><ul><ul><li>ID gene fragments from genomics database </li></ul></ul><ul><ul><li>Tissue distribution of oyster </li></ul></ul><ul><ul><li>Isolate complete ORF or oyster COX gene </li></ul></ul><ul><li>Challenge Experiments </li></ul><ul><li>Applied research </li></ul>
  13. 13. Experiments <ul><li>Vibrio tubiashii exposures </li></ul><ul><li>whole organism plated hemocytes </li></ul>
  14. 14. Analytical Results <ul><li>PGE (ELISA) </li></ul><ul><li>Receptor (PCR) </li></ul>PGE receptor
  15. 15. Vibrio exposure-ELISA results PGE conjugate PGE sample 100% 0%
  16. 16. Gill Tissue/Vibrio– qPCR PGE receptor
  17. 17. Plated hemocytes/Vibrio - qPCR
  18. 18. Outline <ul><li>What are prostaglandins? </li></ul><ul><li>Characterization of genes involved in biosynthesis of prostaglandins </li></ul><ul><li>Experiments and Results </li></ul><ul><li>Applied research </li></ul>
  19. 19. Applied Research <ul><li>Physiological Processes: </li></ul><ul><li>Immune Response </li></ul><ul><li>Ion transport </li></ul><ul><li>Reproduction </li></ul><ul><li>Homeostasis </li></ul>temperature pathogen exposure acidification pesticides
  20. 20. Applied Research <ul><li>Prostaglandins appear to play a role in the immune response of oysters. </li></ul><ul><li>In vitro hemocyte cultures may be a good experimental model to study physiological responses to disruptions of this pathway </li></ul><ul><ul><li>how would changes in environmental conditions (pollutants, temperature, pH) affect the regulation of arachidonic acid metabolism? </li></ul></ul><ul><ul><li>What physiological effects might this have? </li></ul></ul>
  21. 21. Acknowledgements <ul><li>Advisor, Steven Roberts </li></ul><ul><li>Research scientist, Sam White </li></ul><ul><li>Friedman and Seeb Lab </li></ul>

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