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Fulkerson Capstone


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Fulkerson Capstone

  1. 1. A Comparison of Growth and Gene Expression in Two Species of Oysters By: Katie Fulkerson Advisor: Steven Roberts By: Katie Fulkerson Advisor: Steven Roberts
  2. 2. Oyster History <ul><li>Oyster trade began in America in 1608 </li></ul><ul><li>Habitat degradation and overharvest lead to population declines </li></ul><ul><li>Oyster culture seen as solution to saving natural stocks while continuing to meet consumer demand </li></ul>
  3. 3. Cultivated Oysters <ul><li>Atlantic oyster ( Crassostrea virginica ) </li></ul><ul><li>Kumamoto oyster ( Crassostrea sikamea ) </li></ul><ul><li>European flat oyster ( Ostrea edulis ) </li></ul><ul><li>Pacific oyster ( Crassostrea gigas ) </li></ul><ul><li>Olympia oyster ( Ostrea conchaphila ) </li></ul>
  4. 4. Pacific oyster (Crassostrea gigas) : <ul><li>Valuable commercial species </li></ul><ul><li>Imported from Japan </li></ul><ul><li>Growth tends to be rapid when they are young and typically decreases when they reach 4-5 years of age. </li></ul>
  5. 5. Olympia oyster (Ostrea conchaphila) : <ul><li>Native species </li></ul><ul><li>This species experiences a slower growth rate, taking 4-5 years to reach market size (50mm). </li></ul>
  6. 6. Growth is dependent on … <ul><li>Genetics </li></ul><ul><li>Environmental factors </li></ul><ul><ul><li>Water temperature </li></ul></ul><ul><ul><li>Food availability </li></ul></ul><ul><ul><li>Placement in the water column </li></ul></ul><ul><ul><li>Sediment type </li></ul></ul><ul><ul><li>Density of the bed </li></ul></ul>
  7. 7. Objectives <ul><li>Compare growth rates in two species of oysters from the same environment. </li></ul><ul><li>Identify genes that are likely involved in growth in Pacific and Olympia oysters. </li></ul><ul><li>Compare gene expression patterns from Pacific and Olympia tissues extracted during two periods of development. </li></ul>
  8. 8. <ul><li>Oysters were grown in Agate Pass in Kitsap County, WA. </li></ul><ul><li>Measurements taken from umbo to edge, once a month from August to December 2007. </li></ul><ul><li>Tissues samples from the mantle, gills, and muscle were taken in September and November. </li></ul>Objective 1: Growth Rates
  9. 10. Objective 2: Gene Identification <ul><li>Expressed sequence tags using bioinformatic techniques (KSPI, PKCIP, INSIG-2, P450) </li></ul><ul><li>One previously described gene (mGDF) </li></ul>
  10. 11. Molluscan growth differential factor (mGDF) <ul><li>associated with the transforming growth factor beta family of proteins. </li></ul><ul><li>detected in the muscle of O. conchaphila and all tissues from C. gigas . </li></ul>m g ma L C. gigas m g ma O. conchaphila L
  11. 12. Kazal-type serine peptidase inhibitor domain 1 (KSPI) <ul><li>A Serine proteinase inhibitor found in blood plasma, saliva, secretions of pancreas, seminal vesicles, and submandibular glands. </li></ul><ul><li>detected in all tissues samples of both O. conchaphila and C . gigas. </li></ul>m g ma C. gigas L m g ma O. conchaphila L
  12. 13. Protein kinase C inhibitor protein 1 (PKCIP) <ul><li>part of a family of conserved regulator proteins </li></ul><ul><li>plays regulatory roles in multiple cellular processes including differentiation, cell growth, secretion, and muscle contraction. </li></ul><ul><li>detected in the gill and muscle of O. conchaphila and all tissues of C. gigas . </li></ul>m g ma C. gigas L m g ma O. conchaphila L
  13. 14. Insulin-induced gene 2 protein (INSIG-2) <ul><li>involved in metabolic activity, gene transcription, and cell growth. </li></ul><ul><li>not detected in any tissue of either species. </li></ul>m g ma C. gigas L m g ma O. conchaphila L
  14. 15. Cytochrome P450 17-hydroxylase/lyase (P450) <ul><li>a monooxygenase enzyme </li></ul><ul><li>functions in the metabolism of endogenous compounds such as aromatic hydrocarbons </li></ul><ul><li>In mollusks expression is highest in the digestive gland, but it is also found in blood cells, gills, foot, and gonads. </li></ul><ul><li>detected in the gill and muscle of C. gigas . </li></ul>m g ma C. gigas L m g ma O. conchaphila L
  15. 16. Objective 3: Gene Expression <ul><li>Quantitative RT-PCR methods measured and compared gene expression levels in oyster tissue samples. </li></ul>x x muscle muscle muscle Olympia Gill X mantle mantle mantle Pacific P450 INSIG2 PKCIP KSPI mGDF Species Tissues Sampled
  16. 21. Conclusion <ul><li>Four genes associated with growth were successfully identified in these oysters. </li></ul><ul><li>Characterization of tissue expression patterns </li></ul><ul><li>Quantification of expression level in relation to different periods of development. </li></ul><ul><li>With the exception of KSPI all the genes examined varied in expression levels over time. This may indicate a switch in metabolic processes from the growing season (summer) to fasting season (winter). </li></ul>
  17. 22. Acknowledgements <ul><li>Steven Roberts </li></ul><ul><li>Sam White </li></ul><ul><li>Vivianne Barry and Debbie Kay </li></ul><ul><li>Ken Fulkerson </li></ul>
  18. 23. ??Questions??